Team:Paris/17 August 2009
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- | + | Ligation | |
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- | + | Ligation of pSB1A3 w/ A11 or A13 | |
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- | + | Transformation of BBa_B0034 (AmpR), BBa_E0030(KanR), pSB3C5 (CmR), pINTE3 (AmpR), pTA (AmpR), pINTg3p (AmpR), pINK2 and previous ligation using the "heat choc" protocol | |
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Revision as of 10:02, 17 August 2009
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NoteBook
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Brain work
edit please ^^
Lab work
Vector Dephosphorylation
Dephosphorylation:
pSB1A3 digested by EcoRI/PstI, XbaI/PstI and EcoRI/SpeI have to be dephosphorylated before the ligation process.
Mix : Vector : 30uL
Antarctic Phosphatase : 2uL
Buffer : 3,5 uL
Put the solution at 37°C for 30 min
Add 2 uL of enzyme
37°C for 30 min
65°C for 20 min for heat inactivation
Ligation
Ligation of pSB1A3 w/ A11 or A13
Transformation
Transformation of BBa_B0034 (AmpR), BBa_E0030(KanR), pSB3C5 (CmR), pINTE3 (AmpR), pTA (AmpR), pINTg3p (AmpR), pINK2 and previous ligation using the "heat choc" protocol
To do list
edit please ^^