Team:DTU-Denmark/Notebook/20 August 2013
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===purification gels=== | ===purification gels=== | ||
- | Pooled Nir2 and Nir1 samples that gave bands | + | Pooled Nir2 and Nir1 samples for Morten Noerholm that gave bands. |
* 1 kb ladder | * 1 kb ladder | ||
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* Nir1 | * Nir1 | ||
* 1 kb ladder | * 1 kb ladder | ||
+ | |||
+ | [[File:2013-08-20 puri lin pza21ara nir2 nir1.jpg|600px]] | ||
Second purification | Second purification |
Revision as of 13:58, 21 August 2013
20 August 2013
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Contents |
lab 208
Main purpose
- PCR to linearize pZA21::RFP::araBAD
Who was in the lab
Kristian, Helen, Henrike, Julia
Procedure
PCR to linearize pZA21::RFP::araBAD
- Template: pZA21::RFP::araBAD
- Primers: 55a and 55b
- Program:
temperature | time | cycles |
---|---|---|
98C | 2:00 | - |
98C | 0:10 | 36 |
65C | 1:00 | 36 |
72C | 3:00 | 36 |
72C | 5:00 | - |
10C | hold | - |
The table shows the composition of each reaction:
component (per reaction) | 1 | 2 | 3 | 4 | 5 | 6 | Neg |
---|---|---|---|---|---|---|---|
dNTPs | 1uL | 1uL | 1uL | 1uL | 1uL | 1uL | - |
buffer (GC/HF + volume) | HF + 10uL | HF + 10uL | HF + 10uL | GC + 10uL | HF + 10uL | HF + 10uL | HF + 10uL |
X7 polymerase | 0.5uL | 0.5uL | 0.5uL | 0.5uL | 0.5uL | 0.5uL | 0.5uL |
MilliQ water | 29uL | 21.5uL | 31.5uL | 29uL | 31.5uL | 31.5uL | 32.5uL |
template | 1uL | 1uL | 1uL | 1uL | 1uL | 1uL | - |
FW primer 55a | 3uL | 3uL | 3uL | 3uL | 3uL | 3uL | 3uL |
RV primer 55b | 3uL | 3uL | 3uL | 3uL | 3uL | 3uL | 3uL |
DMSO (100%) | 2.5uL (5%) | - | - | 2.5uL (5%) | - | - | - |
Betaine (5M) | - | 10uL (1M) | - | - | - | - | - |
Repeated reaction mix number 1 and ran on same program to get more backbone.
Results
Gels
loaded more samples from the screening PCR for BioBrick creation yesterday
- 1 kb ladder
- screening D12
- screening D11
- screening D10
- screening D9
- screening D8
- screening D7
- screening D6
- screening D5
- screening D4
- screening D3
- screening D2
- screening D1
- screening C12
- screening C11
- screening C10
- screening C9
- screening C8
- 1 kb ladder
- 1 kb ladder
- screening C7
- screening C6
- screening C5
- screening C4
- screening C3
- screening C2
- screening C1
- screening B12
- screening B11
- screening B10
- screening B9
- screening B8
- screening B7
- 1 kb ladder
purification gels
Pooled Nir2 and Nir1 samples for Morten Noerholm that gave bands.
- 1 kb ladder
- pZA21::RFP::AraBAD
- pZA21::RFP::AraBAD
- pZA21::RFP::AraBAD
- Nir2
- Nir2
- Nir2
- Nir1
- Nir1
- Nir1
- Nir1
- 1 kb ladder
Second purification
- 1 kb ladder
- pZA21::RFP::AraBAD, sample 1 (from todays PCR)
- Nir1
- Nir1
- Nir2
- Nir2
- Nir2
- 1 kb ladder
Conclusion
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