Primer extension
Oxford Dictionary of Biochemistry:
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primer extension
a technique used to map the 5′ ends of mRNA transcripts and thereby identify the exact position of the transcription start site in the sequence of a gene. A labelled antisense oligonucleotide primer is hybridized to mRNA close to the 5′ end and extended by the action of reverse transcriptase. The length of the resulting cDNA reflects the distance between the labelled oligonucleotide and the 5′ end of the mRNA template.
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Primer extension
Primer extension is a technique whereby the 5' ends of RNA or DNA can be mapped.
Primer extension can be used to determine the start site of RNA transcription for a known gene. This technique requires a radiolabelled primer (usually 20 - 50 nucleotides in length) which is complementary to a region near the 3' end of the gene. The primer is allowed to anneal to the RNA and reverse transcriptase is used to synthesize cDNA from the RNA until it reaches the 5' end of the RNA. By running the product on a polyacrylamide gel, it is possible to determine the transcriptional start site, as the length of the sequence on the gel represents the distance from the start site to the radiolabelled primer.
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Oxford Dictionary of Biochemistry
Oxford University Press. Oxford Dictionary of Biochemistry and Molecular Biology © 1997, 2000, 2006 All rights reserved.
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