Check Reagent Supplier Cat. no Primer/Sample ID Memo
Purified PCR product POEM5 & POEM6
ddH2O Invurtogen 10977-015
XbaI NEB 37C, Inactivated at 80C
XhoI NEB 37C, Inactivated at 65C
10x Cutsmart buffer NEB B7204
FasGene PCR clean kit Nippon Genetics FG-91302


1 Reaction preparation
  • Mix the following reagents in one tube (ideally from top to bottom)
Check Reagent Volume (µl) per reaction Final concenration Memo
ddH20 5 -
10x CS buffer 1 1 x
DNA 2 1 µg should be enough
NotI-HF 1 20U/µL
HindII-HF 1 20U/µL
Total (µL) 10

2 Reaction
  • Run the following program on a thermal cycler
STEP Action Time Memo
1 37˚C 1 hour Reaction
2 80˚C 20 mins Inativation
7 4˚C FOREVER Stop reaction.

Sanger sequence sample with right band pattern and proceed to I-SceI/I-CeuI digestion.

Dan Yamamoto-Evans 2019/11/29 12:33 Print at 96%

  • interactome/experiments/20191129.1574998758.txt.gz
  • Last modified: 2019/11/29 12:39
  • by dan