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From the various techniques listed below, which one CANNOT be used to precisely map the transcription start-site of a gene ?
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Eduncle Best Answer
S1 nuclease mapping is a nuclease protection assay using nuclease S1. This technique is used to quantify and map RNA transcripts. It is capable of identifying individual RNAs in a mixture of RNA sample of known sequence. It can particularly map introns and 5’ and 3’ ends of transcribed gene regions. Primer extension is a method typically used to map the 5' end(s) of an RNA, thus defining the transcription start site and providing initial evidence for where the promoter is located within a cloned gene. RACE results in the production of a cDNA copy of the RNA sequence of interest, produced through reverse transcription, followed by PCR amplification of the cDNA copies (RT-PCR). The amplified cDNA copies are then sequenced and, if long enough, should map to a unique genomic region. RACE with high-throughput sequencing was first introduced in 2009 as Deep-RACE to perform mapping of Transcription start sites (TSS) of 17 genes in a single cell-line. However, sequencing the region downstream of promoter cannot be used to map transcription start site.