Primer extension: Difference between revisions

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Primer extension is a technique whereby the 5' ends of RNA or DNA can be mapped.

In this technique, a oligonucleotide primer from a known section of the sequence is annealed to the mRNA. Using this annealed primer as a starting point, reverse transcriptase synthesizes complementary cDNA to the mRNA until it reaches the 5' end. By running the new sequence on polyacrimide gel, it is possible for the experimenter to know the location of the transcription start point (the 5' end), as the length of the sequence on the gel represents the distance from the transcription start point to where the primer was first placed.

References

http://www.biochem.arizona.edu/classes/bioc568/primer_extension.htm

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 '''Primer extension''' is a technique whereby the [[5' end]]s of [[RNA]] or [[DNA]] can be mapped.
-In this technique, we need an [[oligonucleotide]] from a transcribed DNA sequence. This oligonucleotide is annealed to the [[mRNA]]. The oligonucleotide sequence is used as a primer for the [[reverse transcriptase]]. Reverse transcriptase synthesizes one half of [[cDNA]] complementary to the mRNA till the 5' end of mRNA. We can then know the length of the new synthetised cDNA from the oligonucleotide sequence to the transcription start point.
+In this technique, a [[oligonucleotide]] primer from a known section of the sequence is annealed to the [[mRNA]]. Using this annealed primer as a starting point, [[reverse transcriptase]] synthesizes complementary [[cDNA]] to the mRNA until it reaches the 5' end. By running the new sequence on [[polyacrimide gel]], it is possible for the experimenter to know the location of the transcription start point (the 5' end), as the length of the sequence on the gel represents the distance from the transcription start point to where the primer was first placed.
 ==References==