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Team:Wageningen UR/Journal/week7
From 2012.igem.org
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Jeroenbosman (Talk | contribs) (→Lab work) |
Jeroenbosman (Talk | contribs) (→week 7: 11 june - 15 may) |
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| - | = week 7: 11 | + | = week 7: 11 June - 17 June = |
== Office work == | == Office work == | ||
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*Prepare sample for PCR | *Prepare sample for PCR | ||
| - | *Run PCR (98°C for 10 seconds, 55 °C for 30 seconds, 72 °C for 30 seconds, 35 cycles, | + | *Run PCR (98°C for 10 seconds, 55 °C for 30 seconds, 72 °C for 30 seconds, 35 cycles, Pfusion enzyme) |
| + | ! due to unknown reasons the standard program was altered, which resulted into a temperature gradient of the annealing temperature from 52- 58 °C. All the products were in the 58 °C region of the temperature gradient | ||
*Prepare PCR-samples for agarose gel | *Prepare PCR-samples for agarose gel | ||
*Run agarose gel (1% w/v agarose) | *Run agarose gel (1% w/v agarose) | ||
*Check gel | *Check gel | ||
| + | All products amplified well, accept for the CCMV unmodified product. Investigation gives the suspicion that the annealing temperature was to high. Gradient PCR will be performed to find out the right annealing temperature for CCMV. | ||
| - | ''' | + | [WORK IN PROGRESS/ Jeroen) |
| + | |||
| + | '''Testing CCMV protocol''' | ||
Monday: | Monday: | ||
Revision as of 08:34, 20 July 2012
week 7: 11 June - 17 June
Office work
Wiki page
Monday:
- update protocol and journal
Other
Tuesday:
- Preperations made for moving to a larger lab
[Meeting]
written by: Mark
Lab work
Bricking of CCMV and d26-CCMV second try
Monday:
- Prepare sample for PCR
- Run PCR (98°C for 10 seconds, 55 °C for 30 seconds, 72 °C for 30 seconds, 35 cycles, Pfusion enzyme)
! due to unknown reasons the standard program was altered, which resulted into a temperature gradient of the annealing temperature from 52- 58 °C. All the products were in the 58 °C region of the temperature gradient
- Prepare PCR-samples for agarose gel
- Run agarose gel (1% w/v agarose)
- Check gel
All products amplified well, accept for the CCMV unmodified product. Investigation gives the suspicion that the annealing temperature was to high. Gradient PCR will be performed to find out the right annealing temperature for CCMV.
[WORK IN PROGRESS/ Jeroen)
Testing CCMV protocol
Monday:
- Prepare samples for ultracentrifuge according to protocol
- Run ultracentrifuge
written by: Mark
