Primer extension analysis


Primer extension analysis is a method used in transcript analysis to identify the start point of transcription.

The procedure is as follows:

1. Design a primer to hybridise to a known sequence within exon 1 of the transcript. Label the 5' end of the primer.

2. Extend the primer from its 3' end; extension will terminate at the 5' end of the mRNA template

3. The number of nucleotides between the 3' end of the primer and the 5' end of the transcript is unknown (x nt's)

4. Run three lanes in an electrophoresis gel:
- Markers
- Primer
- Extended primer

5. The distance in the gel between the 'primer' band and the 'extended primer' band corresponds to x nts.