August/23 August 2009

From 2009.igem.org

1.transformed check Checked the cell cultures transformed and plated out yesterday (8/22) for colony formation and made an approximate count of the number of colonies.

(ligation sample No.) (no. of colonies)
          9                    1
         10                   100~
         11                   100~

inoculate to iquid medium(5mL + Amp 5uL or Kan 25uL)

2.digestion and ligation

'8/22 digestion sample
↓
gel electrophoresis

UP   : ladder , No.8-1.2.3.4.5.  , No.7-1,2,3,4,5,
DOWN : ladder , 1-18L , 1-23J , 1-23L , No.3 , 1-2M , 2-8G

ladder , 1-2M , 1-14H , 1-23L , No.6

gel cut
↓
purification by [QIAquick Nucleotide Removal Kit]
↓
ligation

ligation
DNA        44
10* buffer 5
ligation   1
total      50uL

↓
16°C 3~4hr
↓
transfromation

3.transfromation

ligation sample
⑤⑫⑬⑭⑮
2-18H
1-8E
1-4E
2-16N
2-16L

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