Team:Nevada/Notebook
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==''''''Team Nevada Notebook''''''== | ==''''''Team Nevada Notebook''''''== | ||
- | |||
- | + | '''Week of April 11-17:''' | |
- | '' | + | * ''Bryson, Michael, Senny, Tyler'' |
+ | ** Made tobacco cell (NT-1) media in Dr. Shintani's lab | ||
- | ''Week of April | + | '''Week of April 18-24:''' |
- | ''Week of | + | * ''Bryson'' |
+ | ** EcoRI digest of pBIB | ||
+ | ** Made Na acetate buffer | ||
+ | |||
+ | '''Week of April 25-May 1:''' | ||
- | '' | + | * ''Bryson'' |
+ | ** Ran agarose gel of EcoRI digest | ||
- | ''Week of May | + | '''Week of May 2-8:''' |
- | + | '''Week of May 9-15:''' | |
- | '' | + | * ''Bryson'' |
+ | ** Ran 0.8% agarose gel of pBIB post-phenol:chloroform cleanup. | ||
+ | ** Did minipreps on additional pBIB liquid cultures. | ||
- | ''Week of May 30-June'': | + | '''Week of May 16-22:''' |
+ | |||
+ | * ''Bryson'' | ||
+ | ** Klenow reactions of EcoRI digests | ||
+ | ** Phenol:chloroform cleanup of pBIB prior to ligation | ||
+ | ** Blunt-end ligation of klenowed pBIB | ||
+ | |||
+ | * ''Randy Pares and Vidim Gladwell'' | ||
+ | ** Designed primers pBIB-RB-F, pBIB-RB-R, NOS 3'-Foward Nos3' reverse for sequencing the pBIB plant plasmid. | ||
+ | |||
+ | '''Week of May 23-29:''' | ||
+ | |||
+ | * ''Bryson'' | ||
+ | ** Incubated 50 mL liquid culture of ''E. coli'' with pBIB (samples 3). | ||
+ | ** Miniprepped sample 3, using modified protocol for large/low-copy plasmids. | ||
+ | ** EcoRI digest of uncut sample 3 | ||
+ | ** Prepared 5 mM dNTP stock | ||
+ | |||
+ | '''Week of May 30-June 5:''' | ||
+ | |||
+ | * ''Bryson'' | ||
+ | ** HinD3 digest of uncut sample 3 | ||
+ | ** Ran 08% gels of samples 1-5 to verify complete digestion by EcoRI | ||
+ | ** Pooled uncut samples 3, 4 and 5 (pBIB-pool) | ||
+ | ** Recieved 5 µg of pBIB from Randy and Vadim's maxi-prep (pBIB-maxi) | ||
+ | ** EcoRI digest of pBIB-pool and pBIB-maxi | ||
+ | ** Ran 0.8% agarose gel of EcoRI digests | ||
+ | ** Klenow reactions of pBIB-pool and pBIB-maxi | ||
+ | ** Made glycerol stocks of pBIB samples 1-5 | ||
+ | |||
+ | '''Week of June 6-12:''' | ||
+ | |||
+ | * ''Bryson'' | ||
+ | ** Ligation reactions for pBIB-pool and pBIB-maxi | ||
+ | ** Digested pBIB-pool and pBIB-maxi again with EcoRI to linearize any unmodified pBIB | ||
+ | ** Transformed Top-10 Cells with modified pBIB (designated pBIB#) | ||
+ | *** Obtained two colonies after overnight incubation | ||
+ | ** Line-streaked the two colonies (pBIB# 1 and pBIB# 2) on an LB-Kan plate | ||
+ | |||
+ | '''Week of June 13-19:''' | ||
+ | |||
+ | * ''Bryson'' | ||
+ | ** Prepared liquid cultures of pBIB# 1 and pBIB# 2 | ||
+ | ** Miniprepped liquid cultures of pBIB# | ||
+ | ** EcoRI and HinDIII digests of pBIB# | ||
+ | ** Ran agarose gel of pBIB cut and uncut, pBIB# uncut, pBIB# cut with EcoRI and pBIB# cut with HinDIII to confirm the absence of EcoRI site in pBIB# | ||
+ | ** Single-colony streaked pBIB# 2 on a fresh LB-Kan plate | ||
+ | |||
+ | '''Week of June 20-26:''' | ||
+ | |||
+ | * ''Bryson'' | ||
+ | ** Selected 3 colonies from pBIB# 2 (2-1, 2-2, 2-3) and streaked on a fresh LB-Kan plate | ||
+ | ** Started 20 mL liquid cultures of 2-1, 2-2, and 2-3 | ||
+ | ** Miniprepped samples | ||
+ | ** EcoRI and XbaI digests of samples to confirm that the EcoRI was gone and 2-1, 2-2, 2-3 arose from the same colony | ||
+ | ** 0.8% agarose gel of digests | ||
+ | |||
+ | '''Week of June 27-July 3:''' | ||
+ | |||
+ | '''Week of July 4-10:''' | ||
+ | |||
+ | '''Week of July 11-17:''' | ||
+ | |||
+ | '''Week of July 18-24:''' | ||
+ | |||
+ | '''Week of July 25-31:''' | ||
+ | |||
+ | '''Week of August 1-7:''' | ||
+ | |||
+ | '''Week of August 8-14:''' | ||
+ | |||
+ | '''Week of August 15-21:''' | ||
+ | |||
+ | '''Week of August 22-28:''' |
Revision as of 00:36, 22 June 2010
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'Team Nevada Notebook'
Week of April 11-17:
- Bryson, Michael, Senny, Tyler
- Made tobacco cell (NT-1) media in Dr. Shintani's lab
Week of April 18-24:
- Bryson
- EcoRI digest of pBIB
- Made Na acetate buffer
Week of April 25-May 1:
- Bryson
- Ran agarose gel of EcoRI digest
Week of May 2-8:
Week of May 9-15:
- Bryson
- Ran 0.8% agarose gel of pBIB post-phenol:chloroform cleanup.
- Did minipreps on additional pBIB liquid cultures.
Week of May 16-22:
- Bryson
- Klenow reactions of EcoRI digests
- Phenol:chloroform cleanup of pBIB prior to ligation
- Blunt-end ligation of klenowed pBIB
- Randy Pares and Vidim Gladwell
- Designed primers pBIB-RB-F, pBIB-RB-R, NOS 3'-Foward Nos3' reverse for sequencing the pBIB plant plasmid.
Week of May 23-29:
- Bryson
- Incubated 50 mL liquid culture of E. coli with pBIB (samples 3).
- Miniprepped sample 3, using modified protocol for large/low-copy plasmids.
- EcoRI digest of uncut sample 3
- Prepared 5 mM dNTP stock
Week of May 30-June 5:
- Bryson
- HinD3 digest of uncut sample 3
- Ran 08% gels of samples 1-5 to verify complete digestion by EcoRI
- Pooled uncut samples 3, 4 and 5 (pBIB-pool)
- Recieved 5 µg of pBIB from Randy and Vadim's maxi-prep (pBIB-maxi)
- EcoRI digest of pBIB-pool and pBIB-maxi
- Ran 0.8% agarose gel of EcoRI digests
- Klenow reactions of pBIB-pool and pBIB-maxi
- Made glycerol stocks of pBIB samples 1-5
Week of June 6-12:
- Bryson
- Ligation reactions for pBIB-pool and pBIB-maxi
- Digested pBIB-pool and pBIB-maxi again with EcoRI to linearize any unmodified pBIB
- Transformed Top-10 Cells with modified pBIB (designated pBIB#)
- Obtained two colonies after overnight incubation
- Line-streaked the two colonies (pBIB# 1 and pBIB# 2) on an LB-Kan plate
Week of June 13-19:
- Bryson
- Prepared liquid cultures of pBIB# 1 and pBIB# 2
- Miniprepped liquid cultures of pBIB#
- EcoRI and HinDIII digests of pBIB#
- Ran agarose gel of pBIB cut and uncut, pBIB# uncut, pBIB# cut with EcoRI and pBIB# cut with HinDIII to confirm the absence of EcoRI site in pBIB#
- Single-colony streaked pBIB# 2 on a fresh LB-Kan plate
Week of June 20-26:
- Bryson
- Selected 3 colonies from pBIB# 2 (2-1, 2-2, 2-3) and streaked on a fresh LB-Kan plate
- Started 20 mL liquid cultures of 2-1, 2-2, and 2-3
- Miniprepped samples
- EcoRI and XbaI digests of samples to confirm that the EcoRI was gone and 2-1, 2-2, 2-3 arose from the same colony
- 0.8% agarose gel of digests
Week of June 27-July 3:
Week of July 4-10:
Week of July 11-17:
Week of July 18-24:
Week of July 25-31:
Week of August 1-7:
Week of August 8-14:
Week of August 15-21:
Week of August 22-28: