IGEM:MIT/2006/Notebook/2006-6-28: Difference between revisions
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General ligation considerations: 10μL total volume, 50ng of each DNA (but extra insert can't hurt) | General ligation considerations: 10μL total volume, 50ng of each DNA (but extra insert can't hurt) | ||
BSMT Ligation | '''BSMT Ligation''' | ||
*3μL water | *3μL water | ||
*1μL T4 DNA ligase buffer (vortex well before adding, make sure it smells ''strongly'' like wet dog!!) | *1μL T4 DNA ligase buffer (vortex well before adding, make sure it smells ''strongly'' like wet dog!!) | ||
| Line 13: | Line 13: | ||
*2.5μL cut backbone (50ng) | *2.5μL cut backbone (50ng) | ||
Control Ligation (Barry's biobrick part) | '''Control Ligation (Barry's biobrick part)''' | ||
*3μL water | *3μL water | ||
*1μL T4 DNA ligase buffer (vortex well before adding, make sure it smells ''strongly'' like wet dog!!) | *1μL T4 DNA ligase buffer (vortex well before adding, make sure it smells ''strongly'' like wet dog!!) | ||
Revision as of 19:35, 27 June 2006
Ligation
We'll do 2 ligations: one with the cut CHL backbone and our cut BSMT DNA, and another one with the cut CHL backbone and Barry's cut biobrick part.
Note to Andre: the 3 tubes of cut DNA are in the freezer in the neon yellow rack
General ligation considerations: 10μL total volume, 50ng of each DNA (but extra insert can't hurt)
BSMT Ligation
- 3μL water
- 1μL T4 DNA ligase buffer (vortex well before adding, make sure it smells strongly like wet dog!!)
- .5μL T4 DNA ligase enzyme
- 3μL cut BSMT (70ng)
- 2.5μL cut backbone (50ng)
Control Ligation (Barry's biobrick part)
- 3μL water
- 1μL T4 DNA ligase buffer (vortex well before adding, make sure it smells strongly like wet dog!!)
- .5μL T4 DNA ligase enzyme
- 3μL cut Barry's BB part (90ng)
- 2.5μL cut backbone (50ng)
