Toronto/Lab Notebook
From 2007.igem.org
(Difference between revisions)
| Line 30: | Line 30: | ||
* Made o/n of Q04400 (2005) with high [IPTG] (pSB2k3 is high copy number in LacI- environment) | * Made o/n of Q04400 (2005) with high [IPTG] (pSB2k3 is high copy number in LacI- environment) | ||
* Gel Extract: | * Gel Extract: | ||
| - | ** B0034 | + | ** B0034 A, S/P |
| - | ** E0433 | + | ** E0433 A, X/P |
| - | ** I13033 | + | ** I13033 A, S/P |
| - | ** J06801 | + | ** J06801 A, E/X |
| - | ** J31003 | + | ** J31003 A, X/P |
'''TO DO:''' | '''TO DO:''' | ||
Revision as of 04:07, 6 October 2007
Contents |
Oct. 5, 2007
Yusuf, Maria, Talal, Faareha
SUMMARY:
- Miniprep: J06801 (2005), Q04400 (2005), B0034 (2007), J31005 (2007)
- Length Check:
| Part Name | Part | Plasmid |
| B0034 A | O | O |
| B0034 B | O | O |
| J06801 A | O | O |
| J06801 B | O | O |
| J31005 A | O | O |
| J31005 B | O | O |
| Q04400 A | O | O |
| Q04400 B | O | O |
- Made competent cells
- Ligate N1 A + S01003 D = N2 (overnight)
- Made o/n of Q04400 (2005) with high [IPTG] (pSB2k3 is high copy number in LacI- environment)
- Gel Extract:
- B0034 A, S/P
- E0433 A, X/P
- I13033 A, S/P
- J06801 A, E/X
- J31003 A, X/P
TO DO:
- B0015 (2005) didn't transform, try again
- Complete gel extract procedure
- Ligate:
Oct. 4, 2007
Esther, Neha, Rafsan
SUMMARY:
- Found alternate to I1303: ligate terminator (B0015) and Lux pR promoter (R0062) separately
- Transformed R0062 (2007), B0015 (2005)
- Made o/n of DH5a, DH5a-z1, J06801 (2005), Q04400 (2005), B0034 (2007), J31005 (2007)
- Digest and Gel Extract of N1 A with enzymes S/P
- Quantitation: N1 = 16.8 ng/uL, S01003 (2007) = 4.1 ng/uL
TO DO:
- Make o/n: R0062 (2007), B0015 (2005)
- Miniprep and Check Lengths: J06801 (2005), Q04400 (2005), B0034 (2007), J31005 (2007)
- Ligate N1 and S01003 and transform in DH5a
- Make competent cells
Oct. 3, 2007
Yusuf
SUMMARY:
- Transformed: J06801 (2005), Q04400 (2005), B0034 (2007), J31005 (2007)
- Minipreped I130033 AB, E0433 AB, S03140 AB, J04500 AB, J31003 AB, N1 AB
- Made o/n of DH5a and DH5a-z1 (from July 16 plate - ask Seema if this is ok)
- Length Check (O = match, X = does not match):
| Part Name | Part | Plasmid |
| E0433 A | O | O |
| E0433 B | O | O |
| I13033 A | X | O |
| I13033 B | X | O |
| J04500 A | O | O |
| J04500 B | O | O |
| J31003 A | O | O |
| J31003 B | O | O |
| N1 A | O | O |
| N1 B | O | O |
| S03140 A | O | O |
| S03140 B | O | O |
TO DO:
- Make 5 Kan plates
- Make bottle of LB Broth
- Make competent cells
- Make o/n of J06801 (2005), Q04400 (2005), B0034 (2007), J31005 (2007)
- Ligate N1 + S01003
- Find alternate to I13033
Oct. 2, 2007
Natalie
SUMMARY:
- Made o/n (2 each) of: I130033 AB, E0433 AB, S03140 AB, J04500 AB, J31003 AB, N1 AB
- NOTE: N1 = J34100 C + S01640 B + B0015
TO DO:
- Miniprep all above parts and length check
- Transform:
- J06801 (2005) - it's in the source box (this is a larger composite part containing J06501)
- B0034 - also in source box, if not, get it from Registry plates
- Q04400 - re-transform using more DNA to see if we get more colonies and transform 2005/2006 version in the source box
- J31005 - chloramphenicol resistance, just in case we need it
- Continue cataloging and organization (Charles: I'll be dropping by to help out with that)
Oct. 1, 2007
Time: 3:45 PM - 8:30 PM
Yusuf, Irina
Summary:
- made 5 KAN plates
- made 2 AMP+KAN plates
- made 300 mL LB
Tranformation of following parts:
- I13033 -> KAN
- E0433 -> KAN
- S03140 -> AMP
- J04500 -> KAN
- J31003 -> AMP
- (J23100C + S01640B)+ B0015 -> KAN
Used 4uL of DNA for all transformations
- tried to do DNA extraction for J06501 but somebody already extracted it (wasn't in the well)
- couldn't find it in the igem dna box as well !!!
TO DO:
- MP o/n for the above parts
