Team:Groningen/Notebook/21 July 2009

From 2009.igem.org

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(Vectors)
(Vectors)
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===Vectors===
===Vectors===
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Positive control for the P3 vector done with both new and last years primers.
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Positive control for the J61002 vector with BJ23006 done with both new and last years primers.
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Revision as of 13:32, 21 July 2009

Igemhomelogo.png

Wet

GVP Cluster

Transporters

HmtA
F102471 2009-07-21 01hr 30min PCR1GelMislukt.jpg

The band at ~1150 kb was cut out of the gel and used for pcr described yesterday gave poor results. No band of expected size and strong primerdimer bands Therefor we will redesign the cloning. The Forward primer will be modified into 2 part. which will require an additional PCR but increases the chance of getting product.

HmtA-cloningplan.JPG

Metal Accumulation

Vectors

Positive control for the J61002 vector with BJ23006 done with both new and last years primers.

VR-VF2
Component amount
MasterMix NH4 21 uL
F primer 1 uL
R primer 1 uL
P3 vector 0.5 uL
Taq polymerase 1 uL


Primers
Cup nr F primer R primer
1 VF2 iGEMGr09 VR iGEMGr09
2 VF2 iGEMGr09 VR iGEMGr08 Aug
3 VF2 iGEMGr09 VR iGEMGr June
4 VF2 iGEMGr08 VR iGEMGr09
5 VF2 iGEMGr08 VR iGEMGr08 Aug
6 VF2 iGEMGr08 VR iGEMGr June


VR VF2 program
Stage Temperature Time
Denaturing 95° 5 min
Start Cycles 30X
Denaturing 95° 20 sec
Annealing 60° 20 sec
Elongation 72° 1.20 min
End cycles
Final elongation 72° 10 min
Hold Forever

Dry

April
MTWTFSS
    1 2 3 4 5
6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30
May
MTWTFSS
        1 2 3
4 5 6 7 8 9 10
11 12 13 14 15 16 17
18 19 20 21 22 23 24
25 26 27 28 29 30 31
June
MTWTFSS
1 2 3 4 5 6 7
8 9 10 11 12 13 14
15 16 17 18 19 20 21
22 23 24 25 26 27 28
29 30
July
MTWTFSS
    1 2 3 4 5
6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30 31
August
MTWTFSS
          1 2
3 4 5 6 7 8 9
10 11 12 13 14 15 16
17 18 19 20 21 22 23
24 25 26 27 28 29 30
31
September
MTWTFSS
  1 2 3 4 5 6
7 8 9 10 11 12 13
14 15 16 17 18 19 20
21 22 23 24 25 26 27
28 29 30
October
MTWTFSS
      1 2 3 4
5 6 7 8 9 10 11
12 13 14 15 16 17 18
19 20 21 22 23 24 25
26 27 28 29 30 31
November
MTWTFSS
            1
2 3 4 5 6 7 8
9 10 11 12 13 14 15
16 17 18 19 20 21 22
23 24 25 26 27 28 29
30