MOLECULAR BIOLOGY SERVICES:

    cDNA depletion

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Ribosomal cDNA depletion

cat.# CS014

- non-mRNA-derived cDNAs are depleted while rare mRNA-derived cDNAs are unaffected
- Output cDNA samples are suitable for further next generation sequencing
- The method is optimized for both eukaryotic and prokaryotic cDNA preps

cDNA samples prepared from total RNA may contain up to 95% of high-copy DNA molecules mainly derived from rRNA and tRNA. It is often necessary to remove those cDNAs while preserving quantitative ratios of less frequent molecules derived from mRNA.

Evrogen's Duplex-Specific Nuclease (DSN)-based ribosomal cDNA depletion results in essential decrease of concentrations of high-copy cDNAs mainly derived from ribosomal and transport RNAs. After the depletion relative amount of abundant housekeeping genes shows moderate changes while concentration ratios of less frequent mRNA-derived cDNAs remain nearly unaffected. At the same time ribosomal cDNA concentration decreases by more than 100 times.

The ribosomal cDNA depletion service includes cDNA synthesis using random primers, DSN-mediated non-mRNA-derived cDNA removal and further depletion efficacy control by qPCR (option).

The technology is especially recommended for cDNA samples prepared from partially degraded eukaryotic RNA and from non-polyadenylated prokaryotic RNA. The method is optimized for both eukaryotic and prokaryotic cDNA preps. Ribosomal cDNA depletion can facilitate whole transcriptome research including deep sequencing of mRNA (mRNA-seq) and analysis of gene expression profiles. As quantitative ratios between less abundant mRNA-derived cDNAs are preserved, the method is suitable for any quantitative approaches.

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Relative cDNAs amount before (white columns) and after (orange columns) ribosomal cDNA depletion.

(A) Human cDNA sample: 18S – 18S rRNA, 28S – 28S rRNA, ACTB – β-actin, GAPDH – glyceraldehyde 3-phosphate dehydrogenase, TBP – TATA-binding protein; IGFR – immunoglobulin G Fc receptor II; IFN-γR – interferon γ receptor. (B) E. coli cDNA sample: 16S – 16S rRNA; 23S – 23S rRNA; Adk – adenylate kinase; NarP – nitrate/nitrite response regulator protein; TalB – transaldolase.

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