AllPrep DNA/RNA Mini Kit

Protocol tips

Upstream tips	Protocol tips	Downstream 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.	- 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.

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).

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

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.

Manufacturer protocol

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

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing RNA isolation / purification Tissue - Human Stomach using AllPrep DNA/RNA Mini Kit from Qiagen. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.