IGEM:Peking/2007/Count-Conjugation-Notebook/2007-7-20: Difference between revisions
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==double digesting | ==double digesting E0040 and I7100 for ligation and electrophoresis== | ||
* digesting | * digesting E0040 and I7100 with EcoR1 and Pst1, restectively | ||
* for about 12 hours | * for about 12 hours | ||
* from left to right: | * from left to right: | ||
| Line 8: | Line 8: | ||
2-5 I7100 and EcoR1 and Pst1 | 2-5 I7100 and EcoR1 and Pst1 | ||
6 | 6 E0040 plasmid | ||
7-10 | 7-10 E0040 and EcoR1 and Pst1 | ||
11 marker DL200 plus | 11 marker DL200 plus | ||
==Gel | ==Gel Purification== | ||
* extract pSB3k3(I7100) vector and e0040 fragments from gel | * extract pSB3k3(I7100) vector and e0040 fragments from gel | ||
| Line 20: | Line 20: | ||
* from left to right: | * from left to right: | ||
EGFP( | EGFP(E0040) | ||
pSB3k3(I7100) | pSB3k3(I7100) | ||
Revision as of 18:07, 24 July 2007
double digesting E0040 and I7100 for ligation and electrophoresis
- digesting E0040 and I7100 with EcoR1 and Pst1, restectively
- for about 12 hours
- from left to right:
1 I7100 plasmid
2-5 I7100 and EcoR1 and Pst1
6 E0040 plasmid
7-10 E0040 and EcoR1 and Pst1
11 marker DL200 plus
Gel Purification
- extract pSB3k3(I7100) vector and e0040 fragments from gel
electrophoresis test the products of gel extraction
- from left to right:
EGFP(E0040)
pSB3k3(I7100)
Marker DL200 plus
ligation
- ligate the pSB3k3(I7100) and EGFP(E0040) with T4 ligase
mini-prep I7100 and E0040
single digestion test and electrophoresis
- digest e0040 and r0040 with EcoR1, respectively
- for about 3 hours
- from left to right:
1-3 e0040 and EcoR1
4-5 r0040 and EcoR1
6 Marker
- ! there are abnormal bands in the e0040 samples.
another single digestion test
- digest I7100 with Xba1
- digest e0040(another sample) with EcoR1
- electrophoresis results. From left to right:
1 I7100 plasmid
2-4 I7100 and Xba1
5 e0040 plasmid
6-8 e0040 and EcoR1
9 marker DL2000 plus
10 r0040 and EcoR1 (products of former digestion test)
