16 Gauge Blunt End Needles, 10

Blunt end cloning of PCR products

  End repair: Add 5-10 units of T4 DNApol and incubate at 37C for 5 minutes. Make solution to 2.5 M NH4OAc and add 1 volume of 95% ethanol; let sit for 5 minutes at RT and spin at 14K x g for 20 minutes. Wash with 70

Blunt end cloning of PCR produc

End repair: Add 5-10 units of T4 DNApol and incubate at 37C for 5 minutes. Make solution to 2.5 M NH4OAc and add 1 volume of 95% ethanol; let sit for 5 minutes at RT and spin at 14K x g for 20 minutes. Wash with 70

Blunt End Cloning of PCR Products PCR产物平端克隆

kinase. Incubate at 37ºC for 1 hour.4.Ligation reaction: Blunt end ligation is carried out in the presence of 5% PEG 8000. A microgram of Bluescript SK is blunt ended (cut with EcoRV ) and ligated with 10-100 ng of the purified PCR product in a 50μl