Toronto/Lab Notebook
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== Oct. 6, 2007 == | == Oct. 6, 2007 == | ||
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| + | Anam, Tara, Jovan | ||
*Finished gel purification of J06801, J31003, B0034, E0433, I13033 | *Finished gel purification of J06801, J31003, B0034, E0433, I13033 | ||
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* MP and PD length check I13504 AB | * MP and PD length check I13504 AB | ||
* PD, GE and gel purification Q04400 C | * PD, GE and gel purification Q04400 C | ||
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== Oct. 5, 2007 == | == Oct. 5, 2007 == | ||
Revision as of 02:47, 7 October 2007
Contents |
Oct. 6, 2007
Anam, Tara, Jovan
- Finished gel purification of J06801, J31003, B0034, E0433, I13033
- MP for Q04400CD, R0062AB
- PD length check for Q04400 CD, R0062AB
| Part Name | Part | Plasmid |
| R0062 A | X | X |
| R0062 B | X | X |
| Q04400 C | O | O |
| Q04400 D | X | X |
- Quantitation of J06801, J31003, B0034, E0433, I13033
- Nothing showed up for J06801, I13033
- [J31003] = 2.2 ng/uL
- [B0034] = 9.4 ng/uL
- [E0433] = 13.5 ng/uL
- Ligated B0034 + J31003 = E1b (refer to diagram posted up in wet lab area)
- Transformed N1A + S01003D = N2 and its in incubator.
- MP o/n for I13033A (2 copies), J06801A (2 copies), I13504 AB in incubator.
TO DO:
- Transform E1b
- MP I13033A, and J06801A (and make stock if there isn't any in the -80 freezer of this)
- Don't do PD length check on these because it has already been done before and has passed it.
- MP and PD length check I13504 AB
- PD, GE and gel purification Q04400 C
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 R0062 (2007) AB, Q04400 (2005) with high [IPTG] (pSB2k3 is high copy number in LacI- environment)
- Transformed I13504 (2007)
- 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:
- B0034 A + J31003 A
- I13033 A + E0433 A
- (J23100 + S01640) D + J06801 A
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 = J23100 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
