PCR primers for lamda gt10?

Mick Jones mjones at rpms.ac.uk
Tue Nov 14 06:40:29 EST 1995


Dear Geoff

We have PCR amplified cDNA inserts directly from lambda gt10 and gt11 
plaques, and subsequently sequenced them on an ABI 373A.  We analysed 
over 1,000 clones.

The sequence of the primers were:
for lambda gt10
5´-GGCTTATGAGTATTTCTTCCAGGG-3´
5´-TTGAGCAAGTTCAGCCTGGTTAAG-3´
for lambda gt11
5´-TTGACACCAGACCAACTGGTAATG-3´
5´-GGTGGCGACGACTCCTGGAGCCC-3´

The PCR conditions were as follows:

Pre-heat step of 94C for 2.5 min
Then 30 cycles of 94C, 45 sec; 60C, 30 sec; 72C, 3 min;
Primers were used at 500 ng per 50 ul reaction.

Good luck

Mick
_____________________________________________________________________
Mick Jones                      Tel: 081-740-3328 (+44-81-740-3328)
Department of Virology          FAX: 081-743-8331 (+44-81-743-8331)
RPMS,  Du Cane Road              Email: mjones at rpms.ac.uk 
London,  W12 0NN,  UK          URL: 
http://www.rpms.ac.uk/virology/virolhome.html
***********************************************************
"Smoke me a kipper, I'll be back for breakfast."   Ace Rimmer (Red 
Dwarf)
"Crackin' toast, Gromit."    Wallace (The Wrong Trousers)
_____________________________________________________________________





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