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 purified using the All Prep DNA/RNA kit (Qiagen) following the manufacturer’s protocol, and employing the optional on-column DNase I treatment. The quality of each RNA sample was assessed using a Bioanalyzer (Agilent, Santa Clara, CA) with a quality requirement of an integrity number > 8.0 for subsequent sequencing. RNA from two biological replicates was collected for each strain and condition.

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Manufacturer protocol

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

Download manufacturer protocol

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