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PCR Genotyping from the Embryo Yoke Sac188bio精品生物—专注于实验室精品爆款的电商平台 - 蚂蚁淘旗下精选188款生物医学科研用品
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PCR Genotyping from the Embryo Yoke Sac

DNAbuffer:

1XPCRbuffer
0.45%NP-40
0.45%Tween-20
ProteinaseK100mg/ml

PreparetheDNAforPCR:

  1. Add50µlDNAbuffertoaPCRtube
  2. Digwashedyolksacintotubeandspindown
  3. Incubateat50°Cfor30minutes,
  4. Boilthelysatefor5minutes,spindown,
  5. Take5µlforPCR.

PCRreaction(50µl)contains:

1XPCRbuffer
2mMMgCl2
100nMdNTPs

Primers:

a.0.25µM
b.0.05µM
c.0.2µM

a________c400bp(targetingregion)

a________b200bp(wildtype)

2.5unit/100ulTaqDNApolymerase

PCRcycle:

Initialdenaturingby94°Cfor4minutes
28cyclesof94°Cfor1minute
60°Cfor1minute
72°Cfor1minute
Finalextensionby72°Cfor10minutes


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