Team:Lethbridge/Notebook/Lab Work/Group2

From 2011.igem.org

(Difference between revisions)

Revision as of 02:13, 15 September 2011

Week 1 May 2 - 8 2011
Week 2 May 9 - 15 2011
Week 3 May 16 - 22 2011
Ryan vacationing in Whitefish. Having an awesome time.
Week 4 May 23 - 29 2011
First week Ryan begins working full time in the lab. Successfully used PCR to alter prefix and suffix regions to to the Silver standard for the genes xylE and mms6. That way we can fuse signal sequences. On an agarose gel the DNA strands are of expected size for both xylE and mms6 with the fusion standard. After attempting to cut PCR products of xylE and mms6 and cloning into psb1c3 using red/white screening, no white colonies test positive for the product when the plasmids are restricted and resolved on a gel.
Week 5 May 30 - June 5 2011
The problem with the ligation of PCR products into psb1c3 is diagnosed as a result of prefix and suffix restriction sites being too close to the ends of the PCR products. Instead of using red/white colony screening and psb1c3 the polymerase used in the PCR produces blunt ends and therefore blunt end ligation is used for cloning with puc19. Different conditions are used in the blunt end ligation. Since puc19 has to be cut bluntly half the samples have the blunt end restriction enzyme heat killed. Blue/white colony screening has to be used in for puc19 cloning which requires X-Gal and IPTG. Solutions of these are made for the plates. Attempts to assemble miscellaneous parts from synthesis vectors to psb1c3 results in positive testing white colonies on an agarose gel for k331025, an enhanced cyan fluorescent protein with a ribosomal binding site and a c-terminally fused arginine residue tail.
Week 5 May 30 - June 5 2011
Week 6 June 6 - 12
Week 7 June 13 - 19
Week 8 June 20 - 26
Week 9 June 27 - July 3
Week 10 July 4 - 10
Week 11 July 11 - 17
Week 12 July 18 - 24
Week 13 July 25 - 31
Week 14 August 1 - 7
Week 15 August 8 - 14
Week 16 August 15 - 21
Week 17 August 22 - 28
Week 18 August 29 - September 4
Week 19 September 5 - 11
Week 20 September 12 - 18
Week 21 September 19 - 25
Final Half Week September 26 - 28

@@ Line 40: / Line 40: @@
 Ryan vacationing in Whitefish. Having an awesome time. <br>
 Week 4 May 23 - 29 2011 <br>
-First week Ryan begins working full time in the lab. Successfully used PCR to alter prefix and suffix regions to to the Silver standard for the genes xylE and mms6. That way we can fuse signal sequences. On an agarose gel the DNA strands are of expected size for both xylE and mms6 with the fusion standard. <br>
+First week Ryan begins working full time in the lab. Successfully used PCR to alter prefix and suffix regions to to the Silver standard for the genes xylE and mms6. That way we can fuse signal sequences. On an agarose gel the DNA strands are of expected size for both xylE and mms6 with the fusion standard. After attempting to cut PCR products of xylE and mms6 and cloning into psb1c3 using red/white screening, no white colonies test positive for the product when the plasmids are restricted and resolved on a gel.<br>
+Week 5 May 30 - June 5 2011 <br>
+The problem with the ligation of PCR products into psb1c3 is diagnosed as a result of prefix and suffix restriction sites being too close to the ends of the PCR products. Instead of using red/white colony screening and psb1c3 the polymerase used in the PCR produces blunt ends and therefore blunt end ligation is used for cloning with puc19. Different conditions are used in the blunt end ligation. Since puc19 has to be cut bluntly half the samples have the blunt end restriction enzyme heat killed. Blue/white colony screening has to be used in for puc19 cloning which requires X-Gal and IPTG. Solutions of these are made for the plates. Attempts to assemble miscellaneous parts from synthesis vectors to psb1c3 results in positive testing white colonies on an agarose gel for k331025, an enhanced cyan fluorescent protein with a ribosomal binding site and a c-terminally fused arginine residue tail.<br>
 Week 5 May 30 - June 5 2011 <br>
 Week 6 June 6 - 12 <br>