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Team:DTU-Denmark/Notebook/30 August 2013
From 2013.igem.org
30 August 2013
Contents |
lab 208
Main purpose
Who was in the lab
Procedure
PCR to amplify HAO and cycAX USER fragments
Made 6 reactions for each, using either the extraction PCR fragment or the glycerol stock as template
| label | cyc gly D | cyc gly % | cyc gly 1uL D | cyc ext D | cyc ext % | cyc ext 1uL D |
|---|---|---|---|---|---|---|
| template | glycerol stock | glycerol stock | glycerol stock | extraction PCR fragment | extraction PCR fragment | extraction PCR fragment |
| DMSO | 1,5 | - | 1,5 | 1,5 | - | 1,5 |
| MgCl2 | - | - | 1 | - | - | 1 |
| MQ | 1 | 2,5 | - | 1 | 2,5 | - |
| label | HAO gly D | HAO gly % | HAO gly 1uL D | HAO ext D | HAO ext % | HAO ext 1uL D |
|---|---|---|---|---|---|---|
| template | glycerol stock | glycerol stock | glycerol stock | extraction PCR fragment | extraction PCR fragment | extraction PCR fragment |
| DMSO | 1,5 | - | 1,5 | 1,5 | - | 1,5 |
| MgCl2 | - | - | 1 | - | - | 1 |
| MQ | 1 | 2,5 | - | 1 | 2,5 | - |
Program:
| temperature | time | cycles |
|---|---|---|
| 98C | 2:00 | - |
| 98C | 0:20 | 36 |
| annealing temperature | 1:00 | 36 |
| 72C | extension time | 36 |
| 72C | 5:00 | - |
| 10C | hold | - |
HAO - 59C, 3:00
cycAX - 55C, 1:30
Results
Gel
- 1 kb ladder
- psB1C3 for Nir, 2% DMSO
- psB1C3 for Nir, 2% DMSO, 1 uL 50mM MgCl2
- psB1C3 for Nir, 4% DMSO
- psB1C3 for Nir, 4% DMSO, 1 uL 50mM MgCl2
- psB1C3 for Nir, 1M Betaine
- purification of psB1C3 USER fragment to make Biobricks (escaped from well -> Ethanol?)
- purification of HAO fragment (escaped from well -> Ethanol?)
- purification of AMO fragment (escaped from well -> Ethanol?)
- purification of psB1C3 USER fragment to make Biobricks (used more loading buffer)
- purification of HAO fragment (used more loading buffer)
- purification of AMO fragment (used more loading buffer)
- purification of pZA21::ara(tight)
- purification of pZA21::ara(tight) with endings for Nir
- 1 kb ladder
Conclusion
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