cDNA Synthesis from Small Non Coding RNA (sncRNA)

VERTIS specialises in cDNA synthesis from small non-coding RNA (sncRNA), we continually work on developing and optimizing our technology to achieve the highest quality cDNA synthesis.

We are able to prepare cDNA libraries from microRNA using relatively small amount of starting material and from a large range of sources.

Our Synthesis Process for:

miRNA Isolation
• Isolation of total RNA from cells or tissues
• From the total RNA, the RNA species smaller 500 nt are enriched
• Separation of small RNA on a denaturing 12.5% polyacrylamide gel
• Fraction of small RNA with a length of 15 – 30 nt is eluted form the gel

cDNA Synthesis
• The eluted RNA is first poly(A)-tailed in a controlled reaction using poly(A) polymerase
• Ligation of a synthetic RNA adapter to the 5’ – phosphate of the poly(A)-tailed RNA
• First strand synthesis is performed using oligo(dT) – adapter primer and M-MLV-RNase H-reverse transcriptase
• The resulting cDNA is then PCR-amplified in 15-25 cycles and purified by gel fractionation

Sample requirements
Minimum amounts of total RNA = 10 µg
Small RNA (smaller 500 nt) = 1 µg
Tissue = 100 mg and cells = 10 million

For more information or to discuss cDNA synthesis from sncRNA please contact us directly.