Team:Groningen/Notebook/24 July 2009
From 2009.igem.org
(Difference between revisions)
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+ | {| | ||
+ | | | ||
{| | {| | ||
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{| border="1" | {| border="1" | ||
- | |+ '''PCR1 | + | |+ '''PCR1''' |
! Component !! amount | ! Component !! amount | ||
|- | |- | ||
- | ! | + | ! 2x Phusion MM |
- | | | + | | 12.5 uL |
+ | |- | ||
+ | ! MQ | ||
+ | | 9.5 uL | ||
|- | |- | ||
- | ! | + | ! Fw |
| 1 uL | | 1 uL | ||
|- | |- | ||
- | ! | + | ! mut1RC |
| 1 uL | | 1 uL | ||
|- | |- | ||
- | ! | + | ! DNA |
- | | | + | | 1 uL |
- | + | ||
- | + | ||
- | + | ||
|} | |} | ||
|width="10"| | |width="10"| | ||
| | | | ||
- | |||
<!--Tabel 2 hier--> | <!--Tabel 2 hier--> | ||
{| | {| | ||
Line 46: | Line 47: | ||
|- | |- | ||
|Denaturing | |Denaturing | ||
- | | | + | |98° |
- | | | + | |1.30 min |
|- | |- | ||
| | | | ||
Line 54: | Line 55: | ||
|Denaturing | |Denaturing | ||
|98° | |98° | ||
+ | |10 sec | ||
+ | |- | ||
+ | |Annealing | ||
+ | |55° | ||
|20 sec | |20 sec | ||
+ | |- | ||
+ | |Elongation | ||
+ | |72° | ||
+ | |30 sec | ||
+ | |- | ||
+ | | | ||
+ | |End cycles | ||
+ | |- | ||
+ | |Final elongation | ||
+ | |72° | ||
+ | |10 min | ||
+ | |- | ||
+ | |Hold | ||
+ | |4° | ||
+ | |Forever | ||
+ | |} | ||
+ | |width="10"| | ||
+ | | | ||
+ | <!--Tabel 3 hier--> | ||
+ | |} | ||
+ | |||
+ | |||
+ | {| | ||
+ | | | ||
+ | <!--Tabel 1 hier--> | ||
+ | {| border="1" | ||
+ | |+ '''PCR2''' | ||
+ | ! Component !!amount | ||
+ | |- | ||
+ | ! 2x Phusion MM | ||
+ | | 12.5 uL | ||
+ | |- | ||
+ | ! MQ | ||
+ | | 9.5 uL | ||
+ | |- | ||
+ | ! Rev | ||
+ | | 1 uL | ||
+ | |- | ||
+ | ! mut2RC | ||
+ | | 1 uL | ||
+ | |- | ||
+ | ! DNA | ||
+ | | 1 uL | ||
+ | |} | ||
+ | |||
+ | |width="10"| | ||
+ | | | ||
+ | <!--Tabel 2 hier--> | ||
+ | {| | ||
+ | ! PCR2 program | ||
+ | ! Temperature | ||
+ | ! Time | ||
+ | |- | ||
+ | |Denaturing | ||
+ | |98° | ||
+ | |1.30 min | ||
+ | |- | ||
+ | | | ||
+ | |Start Cycles 30X | ||
+ | |- | ||
+ | |Denaturing | ||
+ | |98° | ||
+ | |10 sec | ||
|- | |- | ||
|Annealing | |Annealing | ||
Line 62: | Line 130: | ||
|Elongation | |Elongation | ||
|72° | |72° | ||
- | | | + | |20 sec |
|- | |- | ||
| | | | ||
Line 74: | Line 142: | ||
|4° | |4° | ||
|Forever | |Forever | ||
+ | |} | ||
+ | |width="10"| | ||
+ | | | ||
+ | <!--Tabel 3 hier--> | ||
|} | |} | ||
+ | |||
+ | |||
+ | {| | ||
+ | | | ||
+ | <!--Tabel 1 hier--> | ||
+ | |||
+ | {| border="1" | ||
+ | |+ '''pBAD24''' | ||
+ | ! Component !! amount | ||
+ | |- | ||
+ | ! mm NHSO | ||
+ | | 21 uL | ||
+ | |- | ||
+ | ! 24_F | ||
+ | | 1 uL | ||
+ | |- | ||
+ | ! 24_R | ||
+ | | 1 uL | ||
+ | |- | ||
+ | ! DNA | ||
+ | | 1 uL | ||
+ | |- | ||
+ | ! taq polymerase | ||
+ | | 1 uL | ||
+ | |- | ||
+ | |} | ||
+ | |||
+ | |width="10"| | ||
+ | | | ||
+ | <!--Tabel 2 hier--> | ||
+ | {| | ||
+ | ! PCR pBAD-24 program | ||
+ | ! Temperature | ||
+ | ! Time | ||
+ | |- | ||
+ | |Denaturing | ||
+ | |95° | ||
+ | |2.00 min | ||
+ | |- | ||
+ | | | ||
+ | |Start Cycles 30X | ||
+ | |- | ||
+ | |Denaturing | ||
+ | |95° | ||
+ | |30 sec | ||
+ | |- | ||
+ | |Annealing | ||
+ | |55° | ||
+ | |20 sec | ||
+ | |- | ||
+ | |Elongation | ||
+ | |72° | ||
+ | |2.10 min | ||
+ | |- | ||
+ | | | ||
+ | |End cycles | ||
+ | |- | ||
+ | |Final elongation | ||
+ | |72° | ||
+ | |10 min | ||
+ | |- | ||
+ | |Hold | ||
+ | |4° | ||
+ | |Forever | ||
+ | |} | ||
+ | |width="10"| | ||
+ | | | ||
+ | <!--Tabel 3 hier--> | ||
|} | |} | ||
Revision as of 10:43, 24 July 2009
[http://2009.igem.org/Team:Groningen http://2009.igem.org/wiki/images/f/f1/Igemhomelogo.png]
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Wet
GVP Cluster
Transporters
Below the PCR1.1 and PCR2 of 24/07,
Metal AccumulationVectorsDry
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