IGEM:MIT/2006/Notebook/2006-6-28: Difference between revisions
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'''''Note to Andre: the 3 tubes of cut DNA are in the freezer in the neon yellow rack''''' | '''''Note to Andre: the 3 tubes of cut DNA are in the freezer in the neon yellow rack''''' | ||
#Ligation #1 | *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) | |||
Revision as of 19:33, 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
- 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)
