Team:Lethbridge/Notebook/Lab Work/August
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==August 3/2010== | ==August 3/2010== | ||
(In Lab: HB)<br> | (In Lab: HB)<br> |
Revision as of 01:33, 25 August 2010
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August 3/2010
(In Lab: HB)
Objective: To restrict pBAD, sRBS, mRBS, TetR, dT and pTet for the assembly of pBAD-sRBS-TetR-dT-pTet
Method: Used Restriction of Plasmid DNA protocol.
- A front verctor was made made in sRBS ,mRBS, dT plasmids using EcoRI and Xbal enzymes
- pDNA that was cut out of plasmid for a front vector was pBAD, TetR, dT and pLacI using EcoRI and SpeI enzymes
- A back vector was made in sRBS and mRBS plasmids with PstI and SpeI
- pDNA that was cut out of plasmid for a back vector was TeTR and it was restricted with XbaI and PstI
Construct | pDNA | buffer | Enzymes |
pBAD-sRBS/mRBS | pBAD | Red | EcoRI and SpeI |
pBAD-sRBS/mRBS | sRBS | Orange | XbaI and EcoRI |
pBAD-sRBS/mRBS | mRBS | Orange | XbaI and EcoRII |
sRBS/mRBS-TetR | sRBS | Red | PstI and SpeI |
sRBS/mRBS-TetR | mRBS | Red | PstI and SpeI |
sRBS/mRBS-TetR | TetR | Tango | XbaI and PstI |
TetR-dT | TetR | Red | EcoRI and SpeI |
TetR-dT | dT | Orange | XbaI and EcoRI |
dT-pTet | dT | Red | EcoRI and SpeI |
dT-pTet | pTet | Orange | XbaI and EcoRI |
pLAcI-sRBS/mRBS | pLacI | Red | EcoRI and SpeI |
pLAcI-sRBS/mRBS | sRBS | Orange | XbaI and EcoRI |
pLAcI-sRBS/mRBS | mRBS | Orange | XbaI and EcoRI |
Mms6-PET28(a) | PET28(a) | Orange | NotI |
- For all reactions
- 158 (µL) MilliQ H2O
- 10 (µL) Buffer
- 0.5(µL) of each enzyme
- 10 (µL) pDNA