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Team:DTU-Denmark/Notebook/10 September 2013
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{{:Team:DTU-Denmark/Templates/StartPage|10 September}} | {{:Team:DTU-Denmark/Templates/StartPage|10 September}} | ||
| - | + | Navigate to the [[Team:DTU-Denmark/Notebook/9_September_2013|Previous]] or the [[Team:DTU-Denmark/Notebook/11_September_2013|Next]] Entry | |
=lab 208= | =lab 208= | ||
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<hr/> | <hr/> | ||
==Who was in the lab== | ==Who was in the lab== | ||
<hr/> | <hr/> | ||
| + | Kristian, Henrike | ||
==Procedure== | ==Procedure== | ||
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Redid colony PCR with sequencing primers instead. Two primer pairs to test for the two USER parts. Pair 1: Nir_FW_4_Seq + Nir_RV_5_Seq . Pair 2: Nir_FW_13_Seq + Nir_RV_14_Seq. | Redid colony PCR with sequencing primers instead. Two primer pairs to test for the two USER parts. Pair 1: Nir_FW_4_Seq + Nir_RV_5_Seq . Pair 2: Nir_FW_13_Seq + Nir_RV_14_Seq. | ||
| - | Used Q5 premix. | + | Used Q5 premix. Calculated annealing temperature for pair 1: 60C, pair 2: 64C |
| + | |||
| + | Note: The resulting gel showed a lot of bands that are consistent for all colonies. We repeated the PCR on the next day for one of the colonies with the following annealing temperatures: pair 1: 67C, pair 2: 68C | ||
==Results== | ==Results== | ||
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* 1 kb ladder | * 1 kb ladder | ||
| - | * colony 1, primer pair 1 | + | * Nir colony 1, primer pair 1 |
| - | * colony 1, primer pair 2 | + | * Nir colony 1, primer pair 2 |
| - | * colony 2, primer pair 1 | + | * Nir colony 2, primer pair 1 |
| - | * colony 2, primer pair 2 | + | * Nir colony 2, primer pair 2 |
| - | * colony 3, primer pair 1 | + | * Nir colony 3, primer pair 1 |
| - | * colony 3, primer pair 2 | + | * Nir colony 3, primer pair 2 |
| - | * colony 4, primer pair 1 | + | * Nir colony 4, primer pair 1 |
| - | * colony 5, primer pair 2 | + | * Nir colony 5, primer pair 2 |
| - | * colony 5, primer pair 1 | + | * Nir colony 5, primer pair 1 |
| - | * colony 5, primer pair 2 | + | * Nir colony 5, primer pair 2 |
| - | * colony 6, primer pair 1 | + | * Nir colony 6, primer pair 1 |
| - | * colony 6, primer pair 2 | + | * Nir colony 6, primer pair 2 |
* 1 kb ladder | * 1 kb ladder | ||
| + | |||
| + | [[File:Colony pcr 10. sep..jpg|600px]] | ||
Navigate to the [[Team:DTU-Denmark/Notebook/9_September_2013|Previous]] or the [[Team:DTU-Denmark/Notebook/11_September_2013|Next]] Entry | Navigate to the [[Team:DTU-Denmark/Notebook/9_September_2013|Previous]] or the [[Team:DTU-Denmark/Notebook/11_September_2013|Next]] Entry | ||
{{:Team:DTU-Denmark/Templates/EndPage}} | {{:Team:DTU-Denmark/Templates/EndPage}} | ||
Latest revision as of 12:13, 4 October 2013
10 September
Contents |
lab 208
Who was in the lab
Kristian, Henrike
Procedure
Colony PCR to verify Nir in pZA21::ara-tight
Redid colony PCR with sequencing primers instead. Two primer pairs to test for the two USER parts. Pair 1: Nir_FW_4_Seq + Nir_RV_5_Seq . Pair 2: Nir_FW_13_Seq + Nir_RV_14_Seq.
Used Q5 premix. Calculated annealing temperature for pair 1: 60C, pair 2: 64C
Note: The resulting gel showed a lot of bands that are consistent for all colonies. We repeated the PCR on the next day for one of the colonies with the following annealing temperatures: pair 1: 67C, pair 2: 68C
Results
Gel
- 1 kb ladder
- Nir colony 1, primer pair 1
- Nir colony 1, primer pair 2
- Nir colony 2, primer pair 1
- Nir colony 2, primer pair 2
- Nir colony 3, primer pair 1
- Nir colony 3, primer pair 2
- Nir colony 4, primer pair 1
- Nir colony 5, primer pair 2
- Nir colony 5, primer pair 1
- Nir colony 5, primer pair 2
- Nir colony 6, primer pair 1
- Nir colony 6, primer pair 2
- 1 kb ladder
