Team:HokkaidoU Japan/Notebook/September6
From 2010.igem.org
(Difference between revisions)
(→1-3A由来のpSB1C3と1-5A(RFP reporter)) |
(→1-3A由来のpSB1C3と1-5A(RFP reporter)) |
||
Line 26: | Line 26: | ||
|2.5 uL | |2.5 uL | ||
|- | |- | ||
- | |style="border-top:1px solid # | + | |style="border-top:1px solid #996;"|'''Total''' |
- | |style="border-top:1px solid # | + | |style="border-top:1px solid #996;"|'''50 uL''' |
|} | |} | ||
Line 52: | Line 52: | ||
|2.5 uL | |2.5 uL | ||
|- | |- | ||
- | |style="border-top:1px solid # | + | |style="border-top:1px solid #996;"|'''Total''' |
- | |style="border-top:1px solid # | + | |style="border-top:1px solid #996;"|'''50 uL''' |
|} | |} | ||
Line 78: | Line 78: | ||
|2.5 uL | |2.5 uL | ||
|- | |- | ||
- | |style="border-top:1px solid # | + | |style="border-top:1px solid #996;"|'''Total''' |
- | |style="border-top:1px solid # | + | |style="border-top:1px solid #996;"|'''50 uL''' |
|} | |} | ||
Line 104: | Line 104: | ||
|2.5 uL | |2.5 uL | ||
|- | |- | ||
- | |style="border-top:1px solid # | + | |style="border-top:1px solid #996;"|'''Total''' |
- | |style="border-top:1px solid # | + | |style="border-top:1px solid #996;"|'''50 uL''' |
|} | |} | ||
<div style="clear:both;"></div> | <div style="clear:both;"></div> |
Revision as of 07:08, 20 September 2010
1-3A由来のpSB1C3と1-5A(RFP reporter)
- 制限酵素処理をする前にプライマーなどを確実に除くため,PCR productsのゲル抽出を行った
- 次の組成で制限酵素処理を行った
Reagent | Amount |
---|---|
RFP (1-5A) | 0.75 uL |
DW | 34.3 uL |
0.1% BSA | 5 uL |
10x M Buffer | 5 uL |
EcoR I | 2.5 uL |
Pst I | 2.5 uL |
Total | 50 uL |
Reagent | Amount |
---|---|
pSB1C3 | 0.75 uL |
DW | 34.3 uL |
0.1% BSA | 5 uL |
10x M Buffer | 5 uL |
EcoR I | 2.5 uL |
Pst I | 2.5 uL |
Total | 50 uL |
Reagent | Amount |
---|---|
RFP (1-5A) | 0.75 uL |
DW | 34.3 uL |
0.1% BSA | 5 uL |
10x H Buffer | 5 uL |
EcoR I | 2.5 uL |
Pst I | 2.5 uL |
Total | 50 uL |
Reagent | Amount |
---|---|
pSB1C3 | 0.75 uL |
DW | 34.3 uL |
0.1% BSA | 5 uL |
10x H Buffer | 5 uL |
EcoR I | 2.5 uL |
Pst I | 2.5 uL |
Total | 50 uL |
- これらをゲル抽し,25 uLのDNase free water
- この時点でそれぞれ22.56 ng/uL,45.12 ng/uLとなっていると見積もられる
- pSB1C3 2 uL, RFP 3 uLにMighty Mix 5 uLとT4 ligase 0.5 uLを加え,Ligationした
- 50 uLのコンピテントセルをつかいTransformationした