Team:KAIT Japan/Notebook
From 2012.igem.org
(Difference between revisions)
Line 24: | Line 24: | ||
*Primer R(10μM) 4μL | *Primer R(10μM) 4μL | ||
*Template(E.coli DH5α) | *Template(E.coli DH5α) | ||
- | + | '''→Reflection:Took colony too many.You should take less colony.'''<br> | |
+ | 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.''' | ||
---- | ---- | ||
Line 32: | Line 40: | ||
#*Marker:pigment(buffer) 1μL,DNA molecule 2μL,TE buffer 3μL | #*Marker:pigment(buffer) 1μL,DNA molecule 2μL,TE buffer 3μL | ||
#*Sample:pigment(buffer) 1μL,sample 5μL | #*Sample:pigment(buffer) 1μL,sample 5μL | ||
- | #*Gel concentration:1.2%,Migration time: | + | #*Gel concentration:1.2%,Migration time:30min |
#Storage | #Storage | ||
===PCR Product 1,3,9=== | ===PCR Product 1,3,9=== |
Revision as of 03:28, 13 August 2012
Home | Team | Project | Parts | Attributions | Notebook | Safety | Sitemap |
---|
During the creation
Creating parts of Tar methylation region.
Contents |
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.
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
- Storage
PCR Product 1,3,9
- Electrophoresis
- The gel check and cut
- DNA purification
- Confirmation of electrophoresis
- PCR
- 50cycle
- Storage