Find the best product for any wet lab experiment Shared knowledge is peer reviewed Ask questions, get answers Toggle navigation Experiments Products Discussions   Login   Join for free Labettor RNA cDNA synthesis Tissue cDNA synthesis Tissue The formation of DNA from an RNA template using reverse transcription, leads to the formation of double stranded complementary DNA, or cDNA. The challenges with this process include: 1. Maintaining the integrity of RNA, 2. Hairpin loops or other secondary structures formed by single stranded RNA can also affect cDNA synthesis, and 3. DNA-RNA hybrids, which may result when the first strand of cDNA is formed. For the first challenge, using workflows that involve proper isolation and storage of RNA, and maintaining a nuclease free environment helps obtain RNA with ideal 260/230 ratios. Using a reverse transcriptase that can tolerate high temperatures (50-55 o C), overcomes obstacles imposed by secondary RNA structures. Finally, RNase H has the ability to hydrolyze RNA before the formation of a second cDNA strand. It is important to ensure that RNase H activity is optimal, because higher RNase H activity leads to premature degradation of RNA template. Many reverse transcriptase offer built-in RNase H activity. shRNA gene silencing 11 Matching solutions 1 Discussion Start discussion Found 1 discussion for this experiment Discussion 1 year ago 1 year ago by Alex Santos Ribeiro cDNA conversion with low concentrations I have extracted RNA from brain tissue but my RNA concentrations are as low as 5ng/ml with my highest being around 80ng/ml. Will I be able to perform cDNA conversion with concentrations as low as these? Comment View 2 comments Share your thoughts or question with experts in your field by adding a discussion! Found 11 matching solutions for this experiment Transcriptor High Fidelity cDNA Synthesis Kit Sigma-Aldrich Upstream tips Protocol tips Downstream tips Denature the template-primer mixture by heating the tube for 10 min at +65°C. Incubate the reaction for 10 to 30 min at +45°C to +55°C. Inactivate Transcriptor High Fidelity Reverse Transcriptase by heating to +85°C for 5 min For downstream PCR, use 1 to 5 μl of the reaction product. The cDNA can be used for amplification without further purification or manipulation (e.g. RNase H-treatment) Protocol tips Denature the template-primer mixture by heating the tube for 10 min at +65°C. Incubate the reaction for 10 to 30 min at +45°C to +55°C. Inactivate Transcriptor High Fidelity Reverse Transcriptase by heating to +85°C for 5 min Downstream tips For downstream PCR, use 1 to 5 μl of the reaction product. The cDNA can be used for amplification without further purification or manipulation (e.g. RNase H-treatment) Manufacturer protocol Publication protocol 1 Relevant paper Tetro cDNA Synthesis Kit Bioline Upstream tips Protocol tips Downstream tips This kit is optimized for RT reactions using a wide range of total RNA amounts (10 pg -2 μg), such that long and low abundance cDNAs can be detected by amplification after cDNA synthesis. Incubate samples at 45 °C for 30 min. Incubate samples at 45 °C for 30 min The use of higher incubation temperatures up to 48 °C may increase the yield of cDNA synthesized in cases of complex RNA secondary structure Upstream tips This kit is optimized for RT reactions using a wide range of total RNA amounts (10 pg -2 μg), such that long and low abundance cDNAs can be detected by amplification after cDNA synthesis. Protocol tips Incubate samples at 45 °C for 30 min. Incubate samples at 45 °C for 30 min Downstream tips The use of higher incubation temperatures up to 48 °C may increase the yield of cDNA synthesized in cases of complex RNA secondary structure Manufacturer protocol Publication protocol 1 Relevant paper SuperScript® II Reverse Transcriptase Thermo Fisher Scientific Upstream tips Protocol tips Downstream tips Heat Oligo mixture to 65°C for 5 min and quick chill on ice and later add reaction components. Mix and incubate at 42°C for 2 min. Add 1 μL (200 units) of SuperScript™ II RT and mix by pipetting gently up and down. Incubate at 42°C for 90 min. Inactivate the reaction by heating at 85°C for 5 min. Protocol tips Heat Oligo mixture to 65°C for 5 min and quick chill on ice and later add reaction components. Mix and incubate at 42°C for 2 min. Add 1 μL (200 units) of SuperScript™ II RT and mix by pipetting gently up and down. Incubate at 42°C for 90 min. Inactivate the reaction by heating at 85°C for 5 min. Manufacturer protocol Publication protocol 1 Relevant paper ReadyScript® cDNA Synthesis Mix Sigma-Aldrich Upstream tips Protocol tips Downstream tips This Kit is optimized for the production of targets < 1kb in length. There will be no loss in functional performance after 20 cycles of freezing on dry ice and thawing on ice For downstream reactions, use 1/5th to 1/10th of the first-strand reaction (2-4 μL) for PCR amplification Upstream tips This Kit is optimized for the production of targets < 1kb in length. There will be no loss in functional performance after 20 cycles of freezing on dry ice and thawing on ice Downstream tips For downstream reactions, use 1/5th to 1/10th of the first-strand reaction (2-4 μL) for PCR amplification Manufacturer protocol Publication protocol 1 Relevant paper ImProm-II™ Reverse Transcription System Promega Upstream tips Protocol tips Downstream tips This kit can be used to reverse transcribe RNA templates starting with either total RNA, poly(A)+ mRNA or synthetic transcript RNA Anneal reaction at 25°C, and incubate for 5 minutes. Extend at 42°C for up to one hour Inactive at 70°C for 15 minutes Upstream tips This kit can be used to reverse transcribe RNA templates starting with either total RNA, poly(A)+ mRNA or synthetic transcript RNA Protocol tips Anneal reaction at 25°C, and incubate for 5 minutes. Extend at 42°C for up to one hour Inactive at 70°C for 15 minutes Manufacturer protocol Publication protocol 1 Relevant paper QuantiTect Reverse Transcription Kit Qiagen Upstream tips Protocol tips Downstream tips For Genomic DNA elimination, incubate reactions for 2 min at 42°C, then place immediately on ice. Add Reverse-transcription reaction components and incubate for 15 min at 42°C. Reverse-transcription reaction components Protocol tips For Genomic DNA elimination, incubate reactions for 2 min at 42°C, then place immediately on ice. Add Reverse-transcription reaction components and incubate for 15 min at 42°C. Reverse-transcription reaction components Manufacturer protocol Publication protocol 1 Relevant paper ProtoScript® II First Strand cDNA Synthesis Kit New England BioLabs Upstream tips Protocol tips Downstream tips Mix RNA sample and primer d(T)23VN and denature RNA for 5 minutes at 70°C. Add M-MuLV Reaction Mixand M-MuLV Enzyme Mix and incubate reaction at 42°C for one hour. Inactivate the enzyme at 80°C for 5 minutes Protocol tips Mix RNA sample and primer d(T)23VN and denature RNA for 5 minutes at 70°C. Add M-MuLV Reaction Mixand M-MuLV Enzyme Mix and incubate reaction at 42°C for one hour. Inactivate the enzyme at 80°C for 5 minutes Manufacturer protocol Publication protocol 1 Relevant paper PrimeScript™ 1st strand cDNA Synthesis Kit Takara Upstream tips Protocol tips Downstream tips After DNAse treatment add Oligo dT Primer,dNTP Mixture,Template RNA and RNase-free dH O and heat at 65°C for 5 min. Add reaction mix to the above mix and incubate at 30°C 10 min* and 42C for 60 min Protocol tips After DNAse treatment add Oligo dT Primer,dNTP Mixture,Template RNA and RNase-free dH O and heat at 65°C for 5 min. Add reaction mix to the above mix and incubate at 30°C 10 min* and 42C for 60 min Manufacturer protocol Publication protocol 1 Relevant paper First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Upstream tips Protocol tips Downstream tips Heat the RNA solution to 65ºC for 10 minutes, then chill on ice. Add first-strand cDNA reaction mix and incubate at 37°C for 1 hour Protocol tips Heat the RNA solution to 65ºC for 10 minutes, then chill on ice. Add first-strand cDNA reaction mix and incubate at 37°C for 1 hour Manufacturer protocol Publication protocol 1 Relevant paper iScript cDNA Synthesis Kit Bio-Rad Laboratories Upstream tips Protocol tips Downstream tips Reverse transcribe for 20 min at 46C For downstream PCR use only 1/10 of the reaction volume (typically 2ul) Protocol tips Reverse transcribe for 20 min at 46C Downstream tips For downstream PCR use only 1/10 of the reaction volume (typically 2ul) Manufacturer protocol Publication protocol 1 Relevant paper qScript cDNA Synthesis Kit Quantabio Upstream tips Protocol tips Downstream tips When required, cDNA product can be diluted with 10 mM Tris-HCl (pH 8.0), 0.1 mM EDTA Protocol tips When required, cDNA product can be diluted with 10 mM Tris-HCl (pH 8.0), 0.1 mM EDTA Manufacturer protocol Publication protocol 1 Relevant paper Can't find the product you've used to perform this experiment? It would be great if you can help us by Adding a product! Become shareholder Discussions About us Contact Privacy Terms

Find the best product for any wet lab experiment Shared knowledge is peer reviewed Ask questions, get answers Toggle navigation Experiments Products Discussions   Login   Join for free Labettor RNA cDNA synthesis Yeast cDNA synthesis Yeast The formation of DNA from an RNA template using reverse transcription, leads to the formation of double stranded complementary DNA, or cDNA. The challenges with this process include: 1. Maintaining the integrity of RNA, 2. Hairpin loops or other secondary structures formed by single stranded RNA can also affect cDNA synthesis, and 3. DNA-RNA hybrids, which may result when the first strand of cDNA is formed. For the first challenge, using workflows that involve proper isolation and storage of RNA, and maintaining a nuclease free environment helps obtain RNA with ideal 260/230 ratios. Using a reverse transcriptase that can tolerate high temperatures (50-55 o C), overcomes obstacles imposed by secondary RNA structures. Finally, RNase H has the ability to hydrolyze RNA before the formation of a second cDNA strand. It is important to ensure that RNase H activity is optimal, because higher RNase H activity leads to premature degradation of RNA template. Many reverse transcriptase offer built-in RNase H activity. shRNA gene silencing 6 Matching solutions Start a discussion Start discussion No discussions found Start your discussion Share your thoughts or question with experts in your field Start a discussion Found 6 matching solutions for this experiment SMARTer® PCR cDNA Synthesis Kit Takara Upstream tips Protocol tips Downstream tips Produces high-quality cDNA from nanograms of total or poly A+ RNA. Store RNA and SMART II A Oligo at –70°C. Store all other reagents at –20°C For ds cDNA Polishing Proteinase K treatment is necessary to inactivate the DNA polymerase activity before proceeding with the ligation steps. Do not chill the tube at –20°C or on ice before centrifuging. Chilling the sample will result in coprecipitation of impurities Upstream tips Produces high-quality cDNA from nanograms of total or poly A+ RNA. Store RNA and SMART II A Oligo at –70°C. Store all other reagents at –20°C Protocol tips For ds cDNA Polishing Proteinase K treatment is necessary to inactivate the DNA polymerase activity before proceeding with the ligation steps. Do not chill the tube at –20°C or on ice before centrifuging. Chilling the sample will result in coprecipitation of impurities Manufacturer protocol Publication protocol 1 Relevant paper Superscript reverse tran-scriptase II (SS RT II) system Thermo Fisher Scientific Upstream tips Protocol tips Downstream tips Heat Oligo mixture to 65°C for 5 min and quick chill on ice and later add reaction components. Mix and incubate at 42°C for 2 min. Add 1 μL (200 units) of SuperScript™ II RT and mix by pipetting gently up and down. Incubate at 42°C for 50 min. Inactivate the reaction by heating at 70°C for 15 min. Protocol tips Heat Oligo mixture to 65°C for 5 min and quick chill on ice and later add reaction components. Mix and incubate at 42°C for 2 min. Add 1 μL (200 units) of SuperScript™ II RT and mix by pipetting gently up and down. Incubate at 42°C for 50 min. Inactivate the reaction by heating at 70°C for 15 min. Manufacturer protocol Publication protocol 1 Relevant paper Transcriptor First Strand cDNA Synthesis Kit Roche Lifesciences Upstream tips Protocol tips Downstream tips Add 0.5 ul (10 U) of Reverse Transcriptase to the reaction mix and incubate for Incubate 30 min at 55°C. Inactivate Transcriptor Reverse Transcriptase by heating to 85°C for 5min Protocol tips Add 0.5 ul (10 U) of Reverse Transcriptase to the reaction mix and incubate for Incubate 30 min at 55°C. Inactivate Transcriptor Reverse Transcriptase by heating to 85°C for 5min Manufacturer protocol Publication protocol 1 Relevant paper ProtoScript® II First Strand cDNA Synthesis Kit New England BioLabs Upstream tips Protocol tips Downstream tips Treat sample with DNase I in the presence of 20 units of RNaseOUT at 37°C for 20 min. Inactivate DNAse with 25 mM EDTA at 80°C for 2 min Upstream tips Treat sample with DNase I in the presence of 20 units of RNaseOUT at 37°C for 20 min. Inactivate DNAse with 25 mM EDTA at 80°C for 2 min Manufacturer protocol Publication protocol 1 Relevant paper First-Strand cDNA Synthesis Kit GE Healthcare Life Sciences Upstream tips Protocol tips Downstream tips Heat the RNA solution to 65ºC for 10 minutes, then chill on ice. Add first-strand cDNA reaction mix and incubate at 37°C for 1 hour Protocol tips Heat the RNA solution to 65ºC for 10 minutes, then chill on ice. Add first-strand cDNA reaction mix and incubate at 37°C for 1 hour Manufacturer protocol Publication protocol 1 Relevant paper iScript cDNA Synthesis Kit Bio-Rad Laboratories Upstream tips Protocol tips Downstream tips Reverse transcribe for 20 min at 46C For downstream PCR use only 1/10 of the reaction volume (typically 2ul) Protocol tips Reverse transcribe for 20 min at 46C Downstream tips For downstream PCR use only 1/10 of the reaction volume (typically 2ul) Manufacturer protocol Publication protocol 1 Relevant paper Can't find the product you've used to perform this experiment? It would be great if you can help us by Adding a product! Become shareholder Discussions About us Contact Privacy Terms

Find the best product for any wet lab experiment Shared knowledge is peer reviewed Ask questions, get answers Toggle navigation Experiments Products Discussions   Login   Join for free Labettor RNA cDNA synthesis Cell lines cDNA synthesis Cell lines The formation of DNA from an RNA template using reverse transcription, leads to the formation of double stranded complementary DNA, or cDNA. The challenges with this process include: 1. Maintaining the integrity of RNA, 2. Hairpin loops or other secondary structures formed by single stranded RNA can also affect cDNA synthesis, and 3. DNA-RNA hybrids, which may result when the first strand of cDNA is formed. For the first challenge, using workflows that involve proper isolation and storage of RNA, and maintaining a nuclease free environment helps obtain RNA with ideal 260/230 ratios. Using a reverse transcriptase that can tolerate high temperatures (50-55 o C), overcomes obstacles imposed by secondary RNA structures. Finally, RNase H has the ability to hydrolyze RNA before the formation of a second cDNA strand. It is important to ensure that RNase H activity is optimal, because higher RNase H activity leads to premature degradation of RNA template. Many reverse transcriptase offer built-in RNase H activity. shRNA gene silencing 9 Matching solutions Start a discussion Start discussion No discussions found Start your discussion Share your thoughts or question with experts in your field Start a discussion Found 9 matching solutions for this experiment RevertAid RT Reverse Transcription Kit Thermo Fisher Scientific Upstream tips Protocol tips Downstream tips The enzyme maintains activity at 42-50 °C and is suitable for synthesis of cDNA up to 13 kb. Use following conditions: 25°C for 5 min, 55°C for 30 min and 75°C for 10 min The reverse transcription reaction product can be directly used in PCR applications or stored at -20 °C for less than one week. For longer storage, -70 °C is recommended Upstream tips The enzyme maintains activity at 42-50 °C and is suitable for synthesis of cDNA up to 13 kb. Protocol tips Use following conditions: 25°C for 5 min, 55°C for 30 min and 75°C for 10 min Downstream tips The reverse transcription reaction product can be directly used in PCR applications or stored at -20 °C for less than one week. For longer storage, -70 °C is recommended Manufacturer protocol Publication protocol 1 Relevant paper SuperScript® II Reverse Transcriptase Thermo Fisher Scientific Upstream tips Protocol tips Downstream tips Heat Oligo mixture to 65°C for 5 min and quick chill on ice and later add reaction components. Mix and incubate at 42°C for 2 min. Add 1 μL (200 units) of SuperScript™ II RT and mix by pipetting gently up and down. Incubate at 42°C for 50 min. Inactivate the reaction by heating at 70°C for 15 min. Protocol tips Heat Oligo mixture to 65°C for 5 min and quick chill on ice and later add reaction components. Mix and incubate at 42°C for 2 min. Add 1 μL (200 units) of SuperScript™ II RT and mix by pipetting gently up and down. Incubate at 42°C for 50 min. Inactivate the reaction by heating at 70°C for 15 min. Manufacturer protocol Publication protocol 1 Relevant paper iScript™ Reverse Transcription Supermix for RT-qPCR Bio-Rad Laboratories Upstream tips Protocol tips Downstream tips Prepare transfection master mix and mix throughly by pippetting. Add 15ul master mix to 5ul of RNA for each RT reaction. Adjust the volume of water if the input of RNA is not to 5 ul input (1ug-1pg). Transcribe for 30 min at 42 °C and the reaction is inactivated for 5 min at 85 °C. Protocol tips Prepare transfection master mix and mix throughly by pippetting. Add 15ul master mix to 5ul of RNA for each RT reaction. Adjust the volume of water if the input of RNA is not to 5 ul input (1ug-1pg). Transcribe for 30 min at 42 °C and the reaction is inactivated for 5 min at 85 °C. Manufacturer protocol Publication protocol 1 Relevant paper QuantiTect Reverse Transcription Kit Qiagen Upstream tips Protocol tips Downstream tips Prepare the reverse-transcription master mix on ice and later incubate for 60 min at 37°C. Incubate for 5 min at 95°C to inactivate Quantiscript Reverse Transcriptase and later use it for downstream experiments Protocol tips Prepare the reverse-transcription master mix on ice and later incubate for 60 min at 37°C. Incubate for 5 min at 95°C to inactivate Quantiscript Reverse Transcriptase and later use it for downstream experiments Manufacturer protocol Publication protocol 1 Relevant paper High-capacity cDNA reverse transcription kit Applied Biosystems Upstream tips Protocol tips Downstream tips This Kit uses the random primer scheme for initiating cDNA synthesis. Add 10 μL of 2X RT master mix and 10 μL of RNA sample and mix. Treat tubes at 25C for 10 minutes, 37C for 2h and 85C for 5 minutes. Upstream tips This Kit uses the random primer scheme for initiating cDNA synthesis. Protocol tips Add 10 μL of 2X RT master mix and 10 μL of RNA sample and mix. Treat tubes at 25C for 10 minutes, 37C for 2h and 85C for 5 minutes. Manufacturer protocol Publication protocol 1 Relevant paper M-MLV Reverse Transcriptase (200 U/µL) Thermo Fisher Scientific Upstream tips Protocol tips Downstream tips Heat mixture to 65°C for 5 minutes and quick chill on ice and incubate at 37°C for 2 minutes. Incubate 50 minutes at 37°C. Inactivate the reaction by heating at 70°C for 15 minutes Protocol tips Heat mixture to 65°C for 5 minutes and quick chill on ice and incubate at 37°C for 2 minutes. Incubate 50 minutes at 37°C. Inactivate the reaction by heating at 70°C for 15 minutes Manufacturer protocol Publication protocol 1 Relevant paper SuperScript IV VILO Master Mix Thermo Fisher Scientific Upstream tips Protocol tips Downstream tips Use up to 2.5 μg of total RNA as starting material in a 20-μL reaction Treat the mixture at 25 °C for 10 min, incubation at 50 °C for 10 min, and incubation at 85 °C for 5 min. Upstream tips Use up to 2.5 μg of total RNA as starting material in a 20-μL reaction Protocol tips Treat the mixture at 25 °C for 10 min, incubation at 50 °C for 10 min, and incubation at 85 °C for 5 min. Manufacturer protocol Publication protocol 1 Relevant paper cDNA SynthePrimeScript™ 1st strand ssynthesis Kit Takara Upstream tips Protocol tips Downstream tips THis kit can be used to synthesize first strand cDNA from total or polyA RNA. Highly sensitive: use less of your precious RNA samples Heat at 65°C for 5 min, then immediately cool on ice Upstream tips THis kit can be used to synthesize first strand cDNA from total or polyA RNA. Highly sensitive: use less of your precious RNA samples Protocol tips Heat at 65°C for 5 min, then immediately cool on ice Manufacturer protocol Publication protocol 1 Relevant paper TaqMan® MicroRNA Reverse Transcription Kit Thermo Fisher Scientific Upstream tips Protocol tips Downstream tips Use 1 to 10 ng of total RNA per 15- µL RT reaction Treat RT mixture for 30 minutes at 16C , 30 minutes at 42C and to inactivate treat the tube at 85C for 5 minutes Upstream tips Use 1 to 10 ng of total RNA per 15- µL RT reaction Protocol tips Treat RT mixture for 30 minutes at 16C , 30 minutes at 42C and to inactivate treat the tube at 85C for 5 minutes Manufacturer protocol Publication protocol 1 Relevant paper Can't find the product you've used to perform this experiment? It would be great if you can help us by Adding a product! Become shareholder Discussions About us Contact Privacy Terms

Find the best product for any wet lab experiment Shared knowledge is peer reviewed Ask questions, get answers Toggle navigation Experiments Products Discussions   Login   Join for free Labettor Discussions Will presence of EDTA effect cDNA synthesis Discussion 8 months ago Stefan Fuhrmann 2 1 Will presence of EDTA effect cDNA synthesis I intend to use iScript cDNA Synthesis Kit in order to synthesize cDNA for qPCR. I have confirmed that my RNA is pure however, according to my extraction protocol I have suspended the RNA in TE buffer containing 1mM EDTA. Will the presence of EDTA have an effect on cDNA synthesis? Experiment: Comment Top comment Answer 1 There is a chance that EDTA might interfere with cDNA synthesis. In order to avoid that you could increase the volume of the cDNA reaction to 50 μl and add extra Magnesium. Make sure that you do not heat the sample in the presence of Magnesium though and only do it before adding it. If this does not work you should use the designated elution buffer of your kit after extracting the RNA instead of TE buffer. Alternatively, you can also use RNAse free water. Hope this helps. Answered 8 months ago Jody Hancock 2 Comments Answer 1 Hi, the EDTA acts as a metal chelator and will inhibit the activity of most enzymes, including reverse transcriptase. I would do a bead clean up and elute your RNA 10 mM TRIS HCL and 50 mM Nacl solution, or something to that end with a neutral pH. You can use the agencourt RNA XP beads for the bead cleanup by beckman coulter. Answered 4 months ago Alex Gomes Answer 1 There is a chance that EDTA might interfere with cDNA synthesis. In order to avoid that you could increase the volume of the cDNA reaction to 50 μl and add extra Magnesium. Make sure that you do not heat the sample in the presence of Magnesium though and only do it before adding it. If this does not work you should use the designated elution buffer of your kit after extracting the RNA instead of TE buffer. Alternatively, you can also use RNAse free water. Hope this helps. Answered 8 months ago Jody Hancock Can you help? Add your comment Start your discussion Share your thoughts or question with experts in your field Start discussion Become shareholder Discussions About us Contact Privacy Terms

Find the best product for any wet lab experiment Shared knowledge is peer reviewed Ask questions, get answers Toggle navigation Experiments Products Discussions   Login   Join for free Labettor RNA RNA isolation Cells primary porcine primary chondrocytes RNA isolation Cells - primary porcine primary chondrocytes When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel. 1 Matching solution 1 Discussion Start discussion Found 1 discussion for this experiment Discussion 1 year ago 1 year ago by Ralf Friedmann I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips? I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips? Comment View 1 comment Share your thoughts or question with experts in your field by adding a discussion! Found 1 matching solution for this experiment PureLink™ RNA Mini Kit Thermo Fisher Scientific Upstream tips Protocol tips Downstream tips - Depending on the material you want to isolate RNA from, please refer to the manual for additional reagents/lab equipment that are required. - DNase is not supplied with this kit, a DNase kit is available which is compatible with this kit. Protocol tips - Depending on the material you want to isolate RNA from, please refer to the manual for additional reagents/lab equipment that are required. Downstream tips - DNase is not supplied with this kit, a DNase kit is available which is compatible with this kit. Manufacturer protocol 1 Relevant paper Can't find the product you've used to perform this experiment? It would be great if you can help us by Adding a product! Become shareholder Discussions About us Contact Privacy Terms

Find the best product for any wet lab experiment Shared knowledge is peer reviewed Ask questions, get answers Toggle navigation Experiments Products Discussions   Login   Join for free Labettor RNA RNA isolation Cells primary human osteoblasts - rheumatoid arthritis RNA isolation Cells - primary human osteoblasts - rheumatoid arthritis When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel. 1 Matching solution 1 Discussion Start discussion Found 1 discussion for this experiment Discussion 1 year ago 1 year ago by Ralf Friedmann I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips? I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips? Comment View 1 comment Share your thoughts or question with experts in your field by adding a discussion! Found 1 matching solution for this experiment RNeasy Plus Mini Kit Qiagen Upstream tips Protocol tips Downstream tips - Include DNAse treatment for 15-20min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C. - Use water to elute the RNA that is warmed to ~60`C. Downstream tips - Include DNAse treatment for 15-20min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C. - Use water to elute the RNA that is warmed to ~60`C. Manufacturer protocol 1 Relevant paper Can't find the product you've used to perform this experiment? It would be great if you can help us by Adding a product! Become shareholder Discussions About us Contact Privacy Terms

Find the best product for any wet lab experiment Shared knowledge is peer reviewed Ask questions, get answers Toggle navigation Experiments Products Discussions   Login   Join for free Labettor RNA RNA isolation Cells primary porcine coronary artery smooth muscle cells RNA isolation Cells - primary porcine coronary artery smooth muscle cells When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel. 1 Matching solution 1 Discussion Start discussion Found 1 discussion for this experiment Discussion 1 year ago 1 year ago by Ralf Friedmann I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips? I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips? Comment View 1 comment Share your thoughts or question with experts in your field by adding a discussion! Found 1 matching solution for this experiment TRIzol Reagent Thermo Fisher Scientific Upstream tips Protocol tips Downstream tips - For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it. Protocol tips - For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it. Manufacturer protocol 1 Relevant paper Can't find the product you've used to perform this experiment? It would be great if you can help us by Adding a product! Become shareholder Discussions About us Contact Privacy Terms

Find the best product for any wet lab experiment Shared knowledge is peer reviewed Ask questions, get answers Toggle navigation Experiments Products Discussions   Login   Join for free Labettor RNA RNA isolation Cells primary bovine coronary artery smooth muscle cells RNA isolation Cells - primary bovine coronary artery smooth muscle cells When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel. 1 Matching solution 1 Discussion Start discussion Found 1 discussion for this experiment Discussion 1 year ago 1 year ago by Ralf Friedmann I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips? I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips? Comment View 1 comment Share your thoughts or question with experts in your field by adding a discussion! Found 1 matching solution for this experiment TRIzol Reagent Thermo Fisher Scientific Upstream tips Protocol tips Downstream tips - For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it. Protocol tips - For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it. Manufacturer protocol 1 Relevant paper Can't find the product you've used to perform this experiment? It would be great if you can help us by Adding a product! Become shareholder Discussions About us Contact Privacy Terms

Find the best product for any wet lab experiment Shared knowledge is peer reviewed Ask questions, get answers Toggle navigation Experiments Products Discussions   Login   Join for free Labettor RNA RNA isolation Cells primary mouse pulmonary artery smooth muscle cells RNA isolation Cells - primary mouse pulmonary artery smooth muscle cells When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel. 1 Matching solution 1 Discussion Start discussion Found 1 discussion for this experiment Discussion 1 year ago 1 year ago by Ralf Friedmann I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips? I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips? Comment View 1 comment Share your thoughts or question with experts in your field by adding a discussion! Found 1 matching solution for this experiment TRIzol Reagent Thermo Fisher Scientific Upstream tips Protocol tips Downstream tips - For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it. Protocol tips - For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it. Manufacturer protocol 1 Relevant paper Can't find the product you've used to perform this experiment? It would be great if you can help us by Adding a product! Become shareholder Discussions About us Contact Privacy Terms

Find the best product for any wet lab experiment Shared knowledge is peer reviewed Ask questions, get answers Toggle navigation Experiments Products Discussions   Login   Join for free Labettor RNA RNA isolation Cells primary rabbit skeletal muscle-derived stem cells RNA isolation Cells - primary rabbit skeletal muscle-derived stem cells When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel. 1 Matching solution 1 Discussion Start discussion Found 1 discussion for this experiment Discussion 1 year ago 1 year ago by Ralf Friedmann I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips? I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips? Comment View 1 comment Share your thoughts or question with experts in your field by adding a discussion! Found 1 matching solution for this experiment TRIzol Reagent Thermo Fisher Scientific Upstream tips Protocol tips Downstream tips - For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it. Protocol tips - For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it. Manufacturer protocol 1 Relevant paper Can't find the product you've used to perform this experiment? It would be great if you can help us by Adding a product! Become shareholder Discussions About us Contact Privacy Terms