IGEM:Peking/2007/Count-Conjugation-Notebook/2007-7-20

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 Extraction

• 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

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)