Next-generation sequencing-based 5 ' rapid amplification of cDNA ends for alternative promoters

作者：Perera Bambarendage P U; Kim Joomyeong^*

来源：Analytical Biochemistry, 2016, 494: 82-84.

DOI：10.1016/j.ab.2015.11.006

• 5 ' RACE
• NGS
• Alternative promoters
• Peg3

Mammalian genomes contain many unknown alternative first exons and promoters. Thus, we have modified the existing 5'RACE (5' rapid amplification of cDNA ends) approach into a next-generation sequencing (NGS)-based new protocol that can identify these alternative promoters. This protocol has incorporated two main ideas: (i) 5'RACE starting from the known second exons of genes and (ii) NGS-based sequencing of the subsequent cDNA products. This protocol also provides a bioinformatics strategy that processes the sequence reads from NGS runs. This protocol has successfully identified several alternative promoters for an imprinted gene, PEG3. Overall, this NGS-based 5'RACE protocol is a sensitive and reliable method for detecting low-abundant transcripts and promoters.

• 出版日期2016-2-1