Material

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



Protocol

1 Reaction preparation
  • Mix the following reagents in one tube (ideally from top to bottom)
Check Reagent Volume (µl) per reaction Final concenration Memo
10x CS buffer 3 1 x
DNA 21 For gel extraction
I-CeuI 3
I-SceI 3
Total (µL) 30



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.



Run gel and extract band.



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

  • interactome/experiments/20191129-2.txt
  • Last modified: 2019/11/29 12:51
  • by dan