AllPrep DNA/RNA Mini Kit

RNA isolation / purification Tissue - Human Stomach

Experiment
RNA isolation / purification Tissue - Human Stomach
Product
AllPrep DNA/RNA Mini Kit from Qiagen
Manufacturer
Qiagen

Protocol tips

Upstream tips
- Complete disruption of plasma membranes of cells and organelles is absolutely required to release all the nucleic acids contained in the sample.

- Different samples require different methods to achieve complete disruption.

- Incomplete disruption results in significantly reduced nucleic acid yields.

- Overloading the spin columns significantly reduces nucleic acid yields.
Protocol tips
- Homogenizing the material is necessary to redice the viscosity of the lysates caused by cell disruption.

- Incomplete homogenization results in inefficient binding of DNA and RNA and therefore significantly reduced yield and purity of nucleic acids.

- Excessive homogenization, on the other hand, results in shorter genomic DNA fragments.

- The centrifugation temperature should be 20–25ºC.

- Warm the lysate to 37ºC before transferring it to the AllPrep DNA spin column.

Publication protocol

Total RNA was extracted after homogenisation with a TissueRuptor rotor–stator using an AllPrep DNA/RNA mini kit (QIAGEN). RNA concentration and purity were assessed using a NanoDrop ND-1000 spectrophotometer (NanoDrop Technologies, DE, USA) and integrity assessed using an Agilent 2100 Bioanalyzer microfluidic platform (Agilent Technologies, CA, USA). After DNase treatment with Ambion Turbo DNA-free kit (Applied Biosystems, CA, USA), cDNA was synthesised using SuperScript II reverse transcriptase with hexamer random primers (both Invitrogen, CA, USA).

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Discussion

Discussion

4 years ago

Author: Paul G. Macon United States

RNA isolation from tissue

How do I extract RNA from animal tissue without using liquid nitrogen? I tried the RNA extraction by using the TRIzol reagent and I homogenize the tissue using polytron homogenizer at room temperature for 30secs is this correct?

Discussion

5 years ago

Author: Aaron Stege Netherlands

Problem in phase separation after using serum/plasma kit

I used a serum/plasma kit for my serum samples. After the phase separation the samples should have 3 phases: a colourless aqueous phase, a white interphase and a red organic phase. However, in some of my samples there was no aqueous phase unless I wait for an extended period of time. How can I circumvent this problem?

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing RNA isolation / purification Tissue - Human Stomach using AllPrep DNA/RNA Mini Kit from Qiagen.

Paper title
Optimising the quantification of cytokines present at low concentrations in small human mucosal tissue samples using Luminex assays
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Manufacturer protocol

Download the product protocol from Qiagen for AllPrep DNA/RNA Mini Kit below.

Download PDF Download manufacturer protocol

Videos

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