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

RNA and DNA were simultaneously extracted from total PBMC or individual immune cell subsets, and purified using the AllPrep DNA/RNA Mini Kit (Qiagen) following the manufacturer’s instruction. RNA quantitation and quality were determined using an Agilent 2100 Bioanalyzer (Agilent and the RNA 6000 Nano Kit (Agilent), according to the manufacturer’s manuals. RNA quality was assessed using the RNA integrity number (RIN), a numerical score of the integrity of RNA, as reported by others [17, 36, 37]. The yield and purity of DNA were determined by spectrophotometric measurements of the ratio of UV absorbance at 260 and 280 nm by a Nanodrop 2000 (Thermo Fisher Scientific).

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Manufacturer protocol

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

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