Team:HokkaidoU Japan/Notebook/August23

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=pSB1C3の活性評価=
=pSB1C3の活性評価=
==EcoR I/Pst I Digestion==
==EcoR I/Pst I Digestion==

Revision as of 14:54, 18 September 2010

Notebook

pSB1C3の活性評価

EcoR I/Pst I Digestion

反応系

pSB1C3 50 uL
DW 4 uL
0.1% BSA 7 uL
10x M Buffer 7 uL
EcoR I 1 uL
Pst I 1 uL
Total 70 uL
  • 37℃，60 min
  • 15 uL 6x Sample Bufferを加え，６レーンに分けて泳動
  • ゲル抽出

Ligationあり

ゲル抽して得た100 uL DNAのうち2 uLを使用してTransformation

Ligationなし

ゲル抽して得た100 uL DNAのうち2 uLに2 uL Ligation Solutionを加える
16℃で30分おき，計4 uLをTransformation

VectorのPCRリベンジ

HokkaidoU Japan 20100823a.JPG

pSB1A3, pSB1C3, pSB1T3それぞれについて以下の反応系でPCRを行った

Vector 1
DW 33
10x Buffer 5
2 M 4dNTPs 5
25 mM MgSO4 3
Suffix-F 1
Prefix-R 1
KOD 1
Total 50 uL
  • 前回と違い，Extentionを120 secで行った
Retrieved from "http://2010.igem.org/Team:HokkaidoU_Japan/Notebook/August23"