PSV质粒扩增
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Lysis buffer：10 mM Tris/HCl (pH 7.8), 10 mM NaCl, 10 mM EDTA,4% SDS, 250 mg proteinase K 21ml？
10ml（about 200ng）psv is incubated with 30ml lysis buffer at 37℃ overnight。
15ml saturated NH4Ac was added to the samples, and was mixed and centrifuged at 13 000 g for 15 min at 4 ℃.
The supernatant was transferred to a new tube and 90 ml cold 99% ethanol was added
DNA was precipitated at 220 uC for 30 min.
Samples were centrifuged at 13 000 g for 5 min at 4 uC and the pellet was washed Tris-EDTA buffer (in-house produced Tris-EDTA).
PCR was performed using AmpliTaq Gold reagents (Roche)
Final concentration of reagents for the 25 ml reaction were 16 PCR buffer