Exo AP

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Description

This is a cheap method to clean PCR product before Sequencing or Genotyping, at 2013 prices it is about 9 cents AUD per reaction. Exonuclease I removes residual single-stranded primers and any extraneous single-stranded DNA produced in the PCR. Antarctic Phosphatase removes the remaining dNTPs from the PCR mixture.

What you need

1. Exonuclease 1

2. Antarctic Phosphatase

both available from Genesearch

Protocol

Reaction mixes for 100 Samples:

Exonuclease 1 (20 U/µl):5µl
Buffer: 10µl
ddH2O: 90µl
Total: 105µl


Antarctic Phosphatase (AP) (5 U/µl): 20µl
Buffer: 10µl
ddH2O: 75µl
Total: 105µl


Make these up in bulk and freeze the two mixes (EXO and AP)

Mix Pools together & Add 2µl per sample (lU each enzyme)

THERMOCYCLER:
37oC for 20 mins
80oC for 10 mins


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