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Team:ITB Indonesia/Notebook/WetLab
From 2013.igem.org
WetLab Notebook
Saturday, 1 June 2013
Making CCMB 80 buffer and CaCl2 + Gliserin 15% buffer.
Sunday, 2 June 2013
Making competent cell with iGEM standard method (CCMB 80 Buffer) and with CaCl2 + Gliserin 15% buffer.
Tuesday, 4 June 2013
Transform the plasmid to DH5α competent cell. We compared number of colony from CaCl2 method and from iGEM standard method. For each method, we spread 20 µl and 200 µl culture.
Wednesday, 5 June 2013
Count the transform colony. Wehave found that the number of colony from 20 µl with CCMB 80 Buffer gave a better result than 200 µl with CaCl2 + Gliserin 15% buffer. So, we are confident with iGEM standard method.
Thursday, 6 June 2013
We try varying the time and repetition of heat shock during transformation. We use single heat shock and double heat shock, and 60 sec and 90 sec for each method.
Friday, 7 June 2013
Count the transformation colony. Wehave found that the number of colony from single heat shock for 60 sec giving the best result than other. So, we applied this method for our transformation.
Saturday, 8 June 2013
Repeating transformation like a day before
Monday, 10 June 2013
- Transform xxx using competent cell and agar plate made of SOB, heat shock 60 seconds 42⁰C, duplo.
Tuesday, 11 june 2013
- Making SOC medium
- Transforming BBa_J04450 (pSB1C3) from transformation eficiency kit (concentration 5pg/ µl) with 1M IPTG induction : 0 µl, 2 µl, 4 µl
- Making 3ml DH5α broth culture from gliserol stock
- Counting the transformacy efficiency from the day before. From first plate resulting 300 colony, and 258 colony from the second
- Discussing 3A assembly system and white screening selection
- Making LB agar stock
Thursday, 13 June 2013
- Making gliserol stock of BBa_J000450. The colonies grew
- Making glycerol stock of BBa_E0430. The colonies grew well.
Friday, June 14th 2013
- Transforming BBa_J22106. The colonies grew poorly
- Transforming BBa_J06702. The colonies grew well.
- Transforming BBa_I13507. The colonies grew well.
- Transforming BBa_I765001. The colonies grew poorly.
- Transforming BBa_E0840. The colonies grew well.
Friday, June 14th 2013
- Preparing solutions for plasmid isolation by alkali lysis method
- Making glycerol stock of 4B2 and 20K3
- Making liquid culture of 9N5, 16B3, 23C3
Monday, June 17th, 2013
- BBa_K592009
- BBa_E1010
- BBa_K592011
- BBa_K592010
Tuesday, June 18th, 2013
- Making liquid culture of 19E1 and 12N3
- Transforming BBa_B0015
- Transforming BBa_B1006
- Making 5X ligation buffer
Wednesday, June 19th, 2013
- Making stock culture of 19E1 and 12N3
- Plasmid isolation from 19E1 and 12N3 stock culture
- Making 1M Tris solution, pH 7.5
- Making 0.5M EDTA slution, pH 8.0
Thursday, June 20th, 2013
- Making stock culture of 4F3 and 6D3
- Plasmid isolation from 4F3 and 6D3 stock culture
- Making and sterilizing TE Buffer
Friday, June 21st, 2013
- Transforming BBa_K873002
- Trannsforming BBa_B0034
Saturday, June 22nd, 2013
- Making glycerol stock of 3H1
- Plasmid isolation from 3H1 stock culture
- Discussing the planned system
Sunday, 23rd, 2013
- Transforming BBa_J23119
Monday, June 24th, 2013
- Confirming results from plasmid isolation by gel electrophoresis
Tuesday, June 25th, 2013
- Plasmid isolation from 18D3 and 18A5 stock culture
Thursday, June 27th, 2013
- Plasmid isolation from 20K3, 16B3, 23C3, 21B5, 9N5, 19E1, 12N3, 4F3, 6D3, and 3HI stock culture
- Confirming results from plasmid isolation in June 25 and 27th by gel electrophoresis
Sunday, June 30th, 2013
- Inoculating (parts?) in 25mL of LB broth + 25 uL of antibiotic
