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Journal of Entomology and Zoology Studies
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P-ISSN: 2349-6800, E-ISSN: 2320-7078

Journal of Entomology and Zoology Studies

2020, Vol. 8, Issue 3
Recent developments in RACE-PCR for the full-length cDNA identification

Samar Jyoti Chutia, Garima Bora, Mukesh Kumar, Rupam Jyoti Nath, Yashwanth BS, Pranab Dihingia, Nishi Sarmah, Nevil Pinto, Prathik MR and Dipak Kumar Sarma

Novel strategies have been developed for the identification of full-length cDNA using bioinformatics tools and multiplexed PCR methods. Usually, sequences are either incomplete or have missing UTR-sequences. Researchers still use the Rapid Amplification of cDNA Ends technique to obtain the full-length cDNA sequences. An oligo(dT) anchor-primer with a hairpin structure is designed for amplification of 3′ cDNA ends. Arbitrary degenerate and sequence-specific reverse primers were also developed for the amplification of 5′ cDNA ends, and tail-PCR is performed until the 5′ sequence of multi-assembled fragment reaches the exon-1 region which can be identified by aligning these fragments to reference genome database. Inhibitory and functional adapters are specially designed; adapter with phosphate can attach to full-length mRNAs with cap structure. In prokaryotes, a technique is reported to capture primary transcripts based on capping the 5′ triphosphorylated RNA. For the identification of full-length cDNA, specific-primers need to be designed for further processes.
Pages : 444-449 | 834 Views | 439 Downloads


Journal of Entomology and Zoology Studies Journal of Entomology and Zoology Studies
How to cite this article:
Samar Jyoti Chutia, Garima Bora, Mukesh Kumar, Rupam Jyoti Nath, Yashwanth BS, Pranab Dihingia, Nishi Sarmah, Nevil Pinto, Prathik MR, Dipak Kumar Sarma. Recent developments in RACE-PCR for the full-length cDNA identification. J Entomol Zool Stud 2020;8(3):444-449.

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