Oligo DNA phosphorylation
Aim
Oligo DNAをアニーリングするときに、vector側をBAP処理すると、Oligoをリン酸化する必要あり
Materials
T4 Polynucleotide kinase (Takara)
10x Polynucleotide kinase Buffer
100 mM ATP
Methods
Phosphorylation
Mixture
50 uM Double strand annealed Oligo DNA 2 ul
T4 Polynucleotide kinase (Takara) 1 ul
10x Polynucleotide kinase Buffer 2 ul
100 mM ATP 0.2 ul
DDW 14.8 ul
total 20 ul
Reaction
37℃ 30 min, 65℃ 15 min, hold 8℃
Dilution
1:50 dilution (e.g DDW 98 uL, phosphorylated dsOligo DNA 2 uL)
Ligation
Remarks
Date : 2018-06-17
Modified Date :
Author : y-goto.icon