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Team:KAIT Japan/Notebook
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#72℃(1min) | #72℃(1min) | ||
#4℃(Save) | #4℃(Save) | ||
| - | #*2~4:35cycle'''→Reflection:The number of cycles was less.''' | + | #*2~4:35cycle'''→Reflection:The number of cycles was less.So,We increased The number of cycles in 8/11.(50 cycles) ''' |
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==Date:8/11== | ==Date:8/11== | ||
===The purified DNA1,2=== | ===The purified DNA1,2=== | ||
Revision as of 16:02, 13 August 2012
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During the creation.
Creating parts of Tar methylation region.
Contents |
Date:8/9
Colony PCR
Reagent
- TaKaRa Ex Taq(5units/μL) 0.5μL
- 10×Ex Taq buffer 10μL
- dNTP Mixture(2.5Meach) 8μL
- Primer F(10μM) 4μL
- Primer R(10μM) 4μL
- Template(E.coli DH5α)
→Reflection:Took colony too many.You should take less colony.
Conditions of the thermal cycler
- 95℃(5min)
- 94℃(30sec)
- 61℃(30sec)
- 71℃(40sec)
- 72℃(1min)
- 4℃(Save)
- 2~4:35cycle→Reflection:The number of cycles was less.So,We increased The number of cycles in 8/11.(50 cycles)
Date:8/11
The purified DNA1,2
- Electrophoresis
- Marker:pigment(buffer) 1μL,DNA molecule 2μL,TE buffer 3μL
- Sample:pigment(buffer) 1μL,sample 5μL
- Gel concentration:1.2%,Migration time:30min
→Reflection:Band was less.
- Storage
PCR Product 1,3,9
- Electrophoresis
- The gel check and cut
- DNA purification
- Confirmation of electrophoresis
- PCR
- 50cycle
- Storage
