D:\533563473.doc 15 March 2007
Enzymatic cleanup of PCR Products
1. Remove excess primers and dNTPs by digesting them
with Exonuclease I (Exo). And shrimp Alkaline
Phosphatase (SAP). Aliquot and mix according to the
following.
2. for 45 l PCR Reaction.
1X
SAP (1 u/l)
1.67 l
EXO I (5u/l)
1.72 l
ddH2O
7.87 l
Total
11.25 l
3.Mix them thoroughly. Add to the PCR Product and
incubate at 370 C for 20 minutes for digestion of primers
and dNTPs,then at 72o C for 20 minutes to kill the enzymes.
The PCR product is not ready for the usual sequencing with
dye primers or dye terminators.
1. SAP from Promega, catalog # M820A 17501505.
2. Exo I from TAKARA ode 2650A lot# K411AA