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Department of Biosciences

Primer Information

 

The service holds the following selection of universal primers in stock.  Simply select a primer appropriate for your application, or alternatively forward a custom primer of your own design.  Custom primers should be supplied at a concentration of 3.2 pmol/μl, a volume of 3μl per reaction is sufficient. Promega have a useful BioMath Calculator on their website for converting primer concentrations to pmol/µl. 

 

 
-21M13 TTg TAA AAC gAC ggC CAg Tg
M13 reverse CAC ACA ggA AAC AgC TAT gAC C
pBluescript SK CgC TCT AgA ACT AgT ggA TC
pBluescript KS CCT CgA ggT CgA Cgg TAT Cg
pBluescript T3 gCA ATT AAC CCT CAC TAA Agg gA
pBluescript/pGEM T7 gTA ATA CgA CTC ACT ATA ggg Cg
pGEM SP6 gCT ATT TAg gTg ACA CTA TAg
pET T7 promoter CgA AAT TAA TAC gAC TCA CTA TAg g
pET T7 terminator ATg CTA gTT ATT gCT CAg Cgg T
Oligo dT TTT TTT TTT TTT TTT TTT TTT TTT Tg/A/C
Oligo dA AAA AAA AAA AAA AAA AAA AAA AAA Ag/T/C

 

Please note that the -21M13 primer held in stock is not compatible with Invitrogen Gateway vectors due to a mismatch at the 3'end. To sequence Gateway vectors please provide the Invitrogen M13(-20) primer or the Stratagene M13(-20) primer.

 

Custom Primer Design

When designing your own primers, please take the following into consideration:

  • Optimum primer length 18-22mer
  • Should have approx. 50-60% GC content
  • Ideally should exhibit GC pairing at both ends
  • Should exhibit nearest GC neighbour interaction, eg. Cg, gC, gg, CC.  Aim for 4-8 pairs, depending on the size of the primer.  A good example is pBluescript KS above, which has 7 pairs
  • Avoid runs of three g bases at 3' end