genepool

Next Generation Sequencing and Bioinformatics
at the University of Edinburgh


Methods used by the GenePool

Sanger Sequencing: SAP Exo I cleaning of PCR products

We use Shrimp Alkaline Phosphatase from Amersham (E70092Z)
and Exonuclease I from New England Biolabs (MO293S).

                      1 sample    1 plate        2 plates      3 plates
   SAP Diln Buffer    1.425 µl    142.5 µl       285 µl        427.5 µl
   SAP (1U)               1 µl      100 µl       200 µl          300 µl
   ExoI (1U)          0.075 µl      7.5 µl        15 µl         22.5 µl

Cycles

   37°C 40 min
   80°C 15 min
   hold at 4°C

1. Make up a master mix and dispense 30 µl into each well of an 8-strip tube.
2. Add 2.5 µl to each well of a plate of PCR products.
3. Seal the plate and place on the PCR machine.


The GenePool has core support from: GenePool admin site
edinburgh
The University of Edinburgh
School of Biological Sciences
The Darwin Trust
of Edinburgh
darwin
mrc
nerc
bbsrc
sulsa

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This is version 2.2 of the GenePool website (July 2009).