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 [http://partsregistry.org/wiki/index.php?title=Part:BBa_K118014 2-18H] [http://partsregistry.org/wiki/index.php?title=Part:BBa_R1051 1-8E] [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0051 1-4E] [http://partsregistry.org/wiki/index.php?title=Part:BBa_K118006 2-16N] [http://partsregistry.org/wiki/index.php?title=Part:BBa_K118005 2-16L]