Get tips on using QIAGEN Large-Construct Kit to perform Plasmid Isolation Clostridium perfringens transconjugants
Get tips on using QIAprep Spin Miniprep Kit to perform Plasmid Isolation Clostridium acetobutylicum/sporogenes
Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Bacteria - Gram positive Clostridum botulinum
Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Bacteria - Gram positive Clostridium tetani
Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Bacteria - Gram positive Clostridium difficile
Get tips on using REzol C&T to perform RNA isolation / purification Bacteria - Gram positive Clostridium difficile
RNA quantification for appropriate concentration and quality (260/280 ratio) is an important step before downstream analysis (including sequencing, RT-qPCR, etc.). Having insufficient RNA quantities or a high salt or phenol in the RNA product can lead to variable or irreproducible downstream results. The various methods used for RNA quantification include: 1. UV spectrophotometric (challenges include: low sensitivity, cannot distinguish between nucleic acid species), 2. Fluorescence-based (challenges include: requires standards, cannot measure amplifiability, not sequence-specific), and 3. RT-PCR (challenges include: requires standards, time-intensive, costly). In order to overcome these challenges, and also to ensure the proper quantification and quality control for RNA product, it is important to use at least two or more methods in order to discard any inconsistencies. Using standards for calibrations increases the sensitivity range for RNA detention (fluorescence- and RT-PCR-based methods). When using RT- PCR, it is important to choose correct primers, aligning to the desired site on the template and of appropriate product length, along with positive, negative and loading controls. It is also important to have at least two primer pairs in order to confirm results.
RNA quantification for appropriate concentration and quality (260/280 ratio) is an important step before downstream analysis (including sequencing, RT-qPCR, etc.). Having insufficient RNA quantities or a high salt or phenol in the RNA product can lead to variable or irreproducible downstream results. The various methods used for RNA quantification include: 1. UV spectrophotometric (challenges include: low sensitivity, cannot distinguish between nucleic acid species), 2. Fluorescence-based (challenges include: requires standards, cannot measure amplifiability, not sequence-specific), and 3. RT-PCR (challenges include: requires standards, time-intensive, costly). In order to overcome these challenges, and also to ensure the proper quantification and quality control for RNA product, it is important to use at least two or more methods in order to discard any inconsistencies. Using standards for calibrations increases the sensitivity range for RNA detention (fluorescence- and RT-PCR-based methods). When using RT- PCR, it is important to choose correct primers, aligning to the desired site on the template and of appropriate product length, along with positive, negative and loading controls. It is also important to have at least two primer pairs in order to confirm results.
Get tips on using NucleoBond® Xtra Midi / Maxi to perform Plasmid Isolation Medicago truncatula BAC clone
Get tips on using QIAamp DNA Mini Kit to perform DNA isolation / purification Bacteria - Gram positive Clostridium difficile
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