DNA + RNA + Protein Extraction Kit
Kit for fractionated isolation of total DNA/RNA/Protein from the same biological sample. Suitable for human and animal tissue, cell culture, plant tissue, bacteria and yeast.
Figure: Analysis of the total RNA fraction isolated with the DNA + RNA + Protein kit. Total RNA preparation from HEP G2 (liver cancer) cell cultures. An optional DNase digestion was not performed. Analysis was performed with an Agilent Bioanalyzer ®.
- Between 100 and 200 ng of high quality total RNA were obtained per column (recommended value: max. 100 ng RNA per column, pictured: 177 ng RNA per column).
- Analysis of triplicate extractions gave RNA integrity numbers (RIN) between 9.50 and 10.00 (pictured: 9.90) for all analyzed samples.
- The rRNA ratio (28S to 18S) was always >2, as expected for high quality RNA preparations (pictured: ~2.5).
- No visual traces of genomic DNA are detectable (if visible, genomic DNA contamination would be detectable as ~ 20 kb sized band). Purified total RNA was free from contaminating DNA, even without any DNase digestion steps.
Results: (1) No product obtained using purified RNA as template; (2) specific product obtained with genomic template DNA as positive control. Positive control DNA template: DNA fraction from the same sample obtained following the DNA purification steps during this extraction. The sensitive PCR assay confirmed the absence of contaminating DNA traces from purified RNA.
Purified total RNA was further used for cDNA synthesis and subsequent qPCR / RealTime PCR analyses with excellent results.
Protocols available for RNA extraction from
- human and animal tissue samples;
- plant tissue samples;
- bacterial cells;
- yeast cells;
- cell culture (human and animal cell lines).
- Simultaneous, fractionated isolation of DNA, RNA and protein from one single sample.
- Protocols available for samples from human or animal tissue, from plant, bacteria and cell cultures.
- Completely phenol-chloroform free purification procedure - no damage or loss of nucleic acids due to attack of free radicals in organic compounds.
- Separates RNA completely from DNA - no DNase digestion required.
- High yield, even with small sample amounts - ideal for scarce sample material (e.g. cell culture monolayers, biopsies).
- dART cDNA Synthesis Kit with all required and specifically adjusted components for Reverse Transcription
- Reverse Transcriptases
- Ribonuclease Inhibitor
- Ribonuclease H ( E.coli ) for selective RNA digestion from DNA/RNA hybrids only
- SG qPCR Master Mixes (2x) for qPCR / RealTime PCR
PCR Master Mix Logsheet