IGEM:Cambridge/2008/Notebook/Bacillus/2008/09/09: Difference between revisions

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- Nanodrop
- Nanodrop
{|class="wikitable" style="text-align:center" border="1"
|-
! product (chromosomal DNA) !! concentration (ng/μL)
|-
| IA751 || 75
|-
| IA751 || ECE153 from 02/09 (on left) || [[Image:photo06/091530203.gif|100px|center]] || colony 1 : zone of clearing, no transformation; colony 2 and 3 : no zone of clearing, transformation ok|| 2 transformed colonies out of 3
|-
| IA751 + ECE112|| 48.6
|-
| IA751 +ECE153 || 74.2
|}


- PCR with AmyE detect primers (12μL of SDW, 5μL of MM, 1+1μL of each primer, 1μL of chromosomal DNA or plasmid)
- PCR with AmyE detect primers (12μL of SDW, 5μL of MM, 1+1μL of each primer, 1μL of chromosomal DNA or plasmid)
- Gel


* Lane1 : HyperladderI
* Lane1 : HyperladderI
Line 34: Line 50:


* Results
* Results
- Only something for IA751, and ECE153 (pure plasmid)???
- New PCR with 4μL of chromosomal DNA and new PCR program
*Result


==PCR AmyE again==
==PCR AmyE again==

Revision as of 08:18, 15 September 2008

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Chromosomal DNA extraction

- ZR soil microbe DNA kit for

  • IA751
  • IA751+ECE112
  • IA751+ECE153

- Nanodrop

product (chromosomal DNA) concentration (ng/μL)
IA751 75
IA751 ECE153 from 02/09 (on left)
File:Photo06/091530203.gif
 colony 1 : zone of clearing, no transformation; colony 2 and 3 : no zone of clearing, transformation ok 2 transformed colonies out of 3
IA751 + ECE112 48.6
IA751 +ECE153 74.2


- PCR with AmyE detect primers (12μL of SDW, 5μL of MM, 1+1μL of each primer, 1μL of chromosomal DNA or plasmid)

- Gel

  • Lane1 : HyperladderI
  • Lane2 : AmyE Back
  • Lane3 : AmyE Front
  • Lane4 : Duncan's DNA
  • Lane5 : ECE112 (plasmid)
  • Lane6 : ECE153 (plasmid)
  • Lane7 : chromosomal DNA IA751 + ECE112
  • Lane8 : chromosomal DNA IA751 + ECE153
  • Lane9 : chromosomal DNA IA751
  • Results

- Only something for IA751, and ECE153 (pure plasmid)???

- New PCR with 4μL of chromosomal DNA and new PCR program

  • Result

PCR AmyE again

- 24μL of SDW, 10μL of MM, 2μL of each primers, 2μL of ECE112

  • Results (cf gel, lane 2 and 3)

- Very good result for amyE front, nothing for AmyE back


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