"
Team:Osaka/week3
From 2012.igem.org
(Difference between revisions)
| (6 intermediate revisions not shown) | |||
| Line 1: | Line 1: | ||
| - | {{ | + | {{Osaka2}} |
__NOTOC__ | __NOTOC__ | ||
| + | |||
==August 12 (Sun)== | ==August 12 (Sun)== | ||
# Miniprep of yesterday's culture | # Miniprep of yesterday's culture | ||
| Line 10: | Line 11: | ||
#* 155(C):1-2D(upstream)+1-23L(downstream)+1-1A(vector) | #* 155(C):1-2D(upstream)+1-23L(downstream)+1-1A(vector) | ||
# SDS-PAGE | # SDS-PAGE | ||
| - | #* | + | #* Electrophoresis |
==August 16 (Thu)== | ==August 16 (Thu)== | ||
# Transformaton of ligation products into DH5α <i>E.coli</i>:153~155 | # Transformaton of ligation products into DH5α <i>E.coli</i>:153~155 | ||
| - | # Transformation of plasmid DNA into Rosetta <i>E.coli</i>: | + | # Transformation of plasmid DNA into Rosetta <i>E.coli</i>:151 |
==August 17 (Fri)== | ==August 17 (Fri)== | ||
| Line 21: | Line 22: | ||
==August 18 (Sat)== | ==August 18 (Sat)== | ||
# Preparation of LB agar plates (Tet,Kan) | # Preparation of LB agar plates (Tet,Kan) | ||
| - | # Miniprep of yesterday's culture | + | # Miniprep of yesterday's culture:153~155 |
# PCR | # PCR | ||
#* mutation was found in PprA → retrial | #* mutation was found in PprA → retrial | ||
# Purification of PCR products | # Purification of PCR products | ||
| + | # Preparation of glycerol stock of cell culture:151 | ||
# Restriction digests of minipreppped parts | # Restriction digests of minipreppped parts | ||
# Gel electrophoresis of digests | # Gel electrophoresis of digests | ||
| Line 30: | Line 32: | ||
# Ligation | # Ligation | ||
#* 156(T):153(upstream)+1-23L(downstream)+1-7A(vector) | #* 156(T):153(upstream)+1-23L(downstream)+1-7A(vector) | ||
| - | # Transformation of ligation products into DH5α E.coli:155,156 | + | # Transformation of ligation products into DH5α <i>E.coli</i>:155,156 |
[[Team:Osaka/Notebook|Back to Notebook]] | [[Team:Osaka/Notebook|Back to Notebook]] | ||
Latest revision as of 01:30, 14 September 2012
August 12 (Sun)
- Miniprep of yesterday's culture
- Restriction digests of minipreppped parts
- Gel electrophoresis of digests
- Ligation
- 153(C):1-2M(upstream)+1-10H(downstream)+1-1A(vector)
- 154(C):1-22A(upstream)+1-2M(downstream)+1-1A(vector)
- 155(C):1-2D(upstream)+1-23L(downstream)+1-1A(vector)
- SDS-PAGE
- Electrophoresis
August 16 (Thu)
- Transformaton of ligation products into DH5α E.coli:153~155
- Transformation of plasmid DNA into Rosetta E.coli:151
August 17 (Fri)
- Transfer to liquid culture
August 18 (Sat)
- Preparation of LB agar plates (Tet,Kan)
- Miniprep of yesterday's culture:153~155
- PCR
- mutation was found in PprA → retrial
- Purification of PCR products
- Preparation of glycerol stock of cell culture:151
- Restriction digests of minipreppped parts
- Gel electrophoresis of digests
- 155 x
- Ligation
- 156(T):153(upstream)+1-23L(downstream)+1-7A(vector)
- Transformation of ligation products into DH5α E.coli:155,156
