Team:Caltech/Week 8

(Difference between revisions)

Revision as of 01:40, 4 August 2010

iGEM 2010

People

Project Details

Protocols

Completed Systems

Notebook

Biosafety

Human Impact

References

Support

Group meeting today!
Started lots more CPCR:
- 2 colonies each of: L24, L22, HSP-Cm, L20, L31, L49, L48, L26, and M30109 using distribution DNA as template.
- Successful: L24-1/-2, 20-4/-5, L22-1/-2, L48-4
Started ligations:
- L49: HSP-Amp + L40 + pSB1T3
- L54: HSP-Amp + K215000 + pSB1K3
- L55: K215000 + I13504 + pSB1K3
Began 2 1mL LB-Kan cultures of 42-4/-5 & incubated @ 30°C
Began 2 5mL LB-Kan cultures of 43-4/-5 & incubated @ 37°C for sequencing tomorrow.

Ligations all failed! (pos control fine)
Prepped 42-4/-5 & sent out for sequencing
Prepped 48-4, 20-4/-5, 22-1/-2, 24-1
- Concentrations all over 200ng/μL, so diluted all 2x with diH2O.
Began colony PCR to verify the contents of the lysis casette and lysis gene (K124017 & K124017)
- After checking the Registry page, we decided that there may be some sort of problem with the DNA or error in its cataloging.
- Also started colony PCR of M30109 cells and ran M30109 digest DNA in the gel to similarly try to verify the presence of the desired insert, after talking with the UNAM-Genomics_Mexico iGEM team.
Digested L23, L44, L22, L24, more Cm and Kan backbone, and a few other bricks
Began ligations:
- L36: K215000 + L22 + pSB1K3
- L38: L19 + L22 + pSB1A3
- L37: L23 + L24 + pSB1K3
- L56: K124017 + B0015 + pSB1C3 (will ligate Thurs)
- L57: K124014 + B0015 + pSB1C3 (will ligate Thurs)

Error creating thumbnail: /websites/2010.igem.org/wiki/bin/ulimit4.sh: line 4: convert: command not found

@@ Line 4: / Line 4: @@
 ==Monday 8/2==
+* Group meeting today!
+* Started lots more CPCR:
+** 2 colonies each of: L24, L22, HSP-Cm, L20, L31, L49, L48, L26, and M30109 using distribution DNA as template.
+** Successful: L24-1/-2, 20-4/-5, L22-1/-2, L48-4
+* Started ligations:
+** L49: HSP-Amp + L40 + pSB1T3
+** L54: HSP-Amp + K215000 + pSB1K3
+** L55: K215000 + I13504 + pSB1K3
+* Began 2 1mL LB-Kan cultures of 42-4/-5 & incubated @ 30&deg;C
+* Began 2 5mL LB-Kan cultures of 43-4/-5 & incubated @ 37&deg;C for sequencing tomorrow.
 ==Tuesday 8/3==
+* Ligations all failed! (pos control fine)
+* Prepped 42-4/-5 & sent out for sequencing
+* Prepped 48-4, 20-4/-5, 22-1/-2, 24-1
+** Concentrations all over 200ng/&mu;L, so diluted all 2x with diH2O.
+* Began colony PCR to verify the contents of the lysis casette and lysis gene (K124017 & K124017)
+** After checking the Registry page, we decided that there may be some sort of problem with the DNA or error in its cataloging.
+** Also started colony PCR of M30109 cells and ran M30109 digest DNA in the gel to similarly try to verify the presence of the desired insert, after talking with the UNAM-Genomics_Mexico iGEM team.
+* Digested L23, L44, L22, L24, more Cm and Kan backbone, and a few other bricks
+* Began ligations:
+** L36: K215000 + L22 + pSB1K3
+** L38: L19 + L22 + pSB1A3
+** L37: L23 + L24 + pSB1K3
+** L56: K124017 + B0015 + pSB1C3 (will ligate Thurs)
+** L57: K124014 + B0015 + pSB1C3 (will ligate Thurs)
 ==Wednesday 8/4==