Difference between revisions of "Team:Technion Israel/Lab Notebook/Overview"
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Revision as of 14:08, 16 August 2015
Lab Notebook Overview
Week 1: May 2-8
Bacillus
We designed gblocks for our gene (3-α-HSD): one for the original sequence and one Bacillus-optimized.
Active team members this week: Liron, Tal, and Ruth
E.coli
Planned the Gibson reaction for our gBlock of 3α-HSD and the plasmid N155 (from our mentors) containing pT7, RBS and mcherry.
Ordered the gBlock of 3α-HSD, which was optimized for E. coli codon usage. We also ordered primers for the Gibson reaction for the gBlock.
Performed reverse PCR to the N155 plasmid and carried out a DPN1 reaction on the product.
Active team members this week: Adi and Alexey
Modeling
content
Comb
content
NADPH
This week we did a lot of research. We found that the 3-α-HSD enzyme can use both NADH and NADPH as co-factors for the reduction reaction in order to inactivate DHT to 3-α-diol.
NADPH has been found to not only enable the reaction in the direction we want, but also to inhibit the reverse reaction if DHT (Rizner, et al.) Therefore, we have decided to try to overproduce NADPH in a host organism, in order to support the activity of the enzyme.
Active team members this week: Yael and Roni
Week 2: May 9-15
Bacillus
We received the gblocks and performed PCR. We had some difficulties amplifying the Bacillus-optimized gblock, and since we found out that B.subtilis has no preferred codon usage, we decided to continue only with the gblock of the original sequence.
Active team members this week: Liron, Tal, and Ruth
E.coli
Our gBlock arrived!!!
We performed PCR on the 3αHSD gBlock with only 10 cycles (as recommended), but received no band in the gel
We performed PCR on the 3αHSD gBlock in higher temperatures, but still received no bands in the gel
Active team members this week: Adi
Modeling
content
Comb
content
NADPH
We conducted further research about the production of NADPH. We sent e-mails to authors of journal articles to enquire about genes and knockouts associated with NADPH overproduction.
Active team members this week: Yael and Roni
Week 3: May 16-22
Bacillus
content
E.coli
Performed PCR on the 3αHSD gBlock in lower temperatures, no bands in the gel.
Active team members this week: Alexey
Modeling
content
Comb
content
NADPH
content
Week 4: May 23-29
Bacillus
content
E.coli
Performed PCR on the 3αHSD gBlock in lower temperatures, no bands in the gel.
Active team members this week: Alexey
Modeling
content
Comb
content
NADPH
content
Week 5: May 30-June 5
Bacillus
content
E.coli
Performed PCR on the 3αHSD gBlock in lower temperatures, no bands in the gel.
Active team members this week: Alexey
Modeling
content
Comb
content
NADPH
content
Week 6: June 6-12
Bacillus
content
E.coli
Performed PCR on the 3αHSD gBlock in lower temperatures, no bands in the gel.
Active team members this week: Alexey
Modeling
content
Comb
content
NADPH
content
Week 7: June 13-19
Bacillus
content
E.coli
Performed PCR on the 3αHSD gBlock in lower temperatures, no bands in the gel.
Active team members this week: Alexey
Modeling
content
Comb
content
NADPH
content
Week 8: June 20-26
Bacillus
content
E.coli
Performed PCR on the 3αHSD gBlock in lower temperatures, no bands in the gel.
Active team members this week: Alexey
Modeling
content
Comb
content
NADPH
content
Week 9: June 27-July 3
Bacillus
content
E.coli
Performed PCR on the 3αHSD gBlock in lower temperatures, no bands in the gel.
Active team members this week: Alexey
Modeling
content
Comb
content
NADPH
content
Week 10: July 4-10
Bacillus
content
E.coli
We took a break for our exams.
Modeling
content
Comb
content
NADPH
content
Week 11: July 11-17
Bacillus
content
E.coli
Performed PCR on the 3αHSD gBlock in lower temperatures, no bands in the gel.
Active team members this week: Alexey
Modeling
content
Comb
content
NADPH
content
Week 12: July 18-24
Bacillus
content
E.coli
Performed PCR on the 3αHSD gBlock in lower temperatures, no bands in the gel.
Active team members this week: Alexey
Modeling
content
Comb
content
NADPH
content
Week 13: July 25-31
Bacillus
content
E.coli
Performed PCR on the 3αHSD gBlock in lower temperatures, no bands in the gel.
Active team members this week: Alexey
Modeling
content
Comb
content
NADPH
content
Week 14: August 1-7
Bacillus
content
E.coli
Performed PCR on the 3αHSD gBlock in lower temperatures, no bands in the gel.
Active team members this week: Alexey
Modeling
content
Comb
content
NADPH
content
Week 15: August 8-14
Bacillus
content
E.coli
Performed PCR on the 3αHSD gBlock in lower temperatures, no bands in the gel.
Active team members this week: Alexey
Modeling
content
Comb
content
NADPH
content
Week 16: August 15-21
Bacillus
content
E.coli
Performed PCR on the 3αHSD gBlock in lower temperatures, no bands in the gel.
Active team members this week: Alexey
Modeling
content
Comb
content
NADPH
content
Week 17: August 22-28
Bacillus
content
E.coli
Performed PCR on the 3αHSD gBlock in lower temperatures, no bands in the gel.
Active team members this week: Alexey
Modeling
content
Comb
content
NADPH
content
Week 18: August 29-September 4
Bacillus
content
E.coli
Performed PCR on the 3αHSD gBlock in lower temperatures, no bands in the gel.
Active team members this week: Alexey
Modeling
content
Comb
content
NADPH
content
Week 19: September 5-11
Bacillus
content
E.coli
Performed PCR on the 3αHSD gBlock in lower temperatures, no bands in the gel.
Active team members this week: Alexey
Modeling
content
Comb
content
NADPH
content
Week 20: September 12-18
Bacillus
content
E.coli
Performed PCR on the 3αHSD gBlock in lower temperatures, no bands in the gel.
Active team members this week: Alexey
Modeling
content
Comb
content
NADPH
content