Article

High-throughput verification of transcriptional starting sites by Deep-RACE

Functional Genomics Technology Team, Omics Science Center, RIKEN Yokohama Institute, Kanagawa, Japan.
BioTechniques (Impact Factor: 2.75). 03/2009; 46(2):130-2. DOI: 10.2144/000113066
Source: PubMed

ABSTRACT We present a high-throughput method for investigating the transcriptional starting sites of genes of interest, which we named Deep-RACE (Deep-rapid amplification of cDNA ends). Taking advantage of the latest sequencing technology, it allows the parallel analysis of multiple genes and is free of time-consuming cloning steps. In comparison to the sequencing of RACE PCR products, our approach is more precise and more cost-effective even for batches as small as 17.

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Available from: Piero Carninci, Aug 23, 2015
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    • "3 0 -RACE-PCR was performed using UPM together with an LvCTL3-specific forward primer 3RACE1, and the nested PCR was subsequently performed with NUP and LvCTL3–3RACE2. The second PCR products were cloned into pMD-20T vector (TaKaRa, Japan) and 12 positive clones were selected and sequenced (ABI PRISM, Applied Biosystems, USA). 5 0 -RACE is a technique used in molecular biology to obtain the 5 0 UTR of a transcript and identify the transcription starting site (TSS) of promoter elements (Olivarius et al., 2009; Scotto-Lavino et al., 2006). TSS of LvCTL3 is determined according to the 5 0 -RACE PCR amplification. "
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