"
Team:WHU-China/Notebook
From 2011.igem.org
(Difference between revisions)
| Line 73: | Line 73: | ||
<div id="daytoday"> | <div id="daytoday"> | ||
<img src="/wiki/images/8/83/Whu-Progress.png"/> | <img src="/wiki/images/8/83/Whu-Progress.png"/> | ||
| - | <img src="/wiki/images/0/02/Whu-Progress.jpg" style="position:absolute;left:20px;top: | + | <img src="/wiki/images/0/02/Whu-Progress.jpg" style="position:absolute;left:20px;top:53px;width:400px;height:320px;"> |
<div style="position:absolute;top:60px;right:30px;width:520px;font-size:15px;"> | <div style="position:absolute;top:60px;right:30px;width:520px;font-size:15px;"> | ||
The forth layer assembly and assembled CHL1-3:</br> | The forth layer assembly and assembled CHL1-3:</br> | ||
Revision as of 09:00, 3 October 2011
• Dealing with biobricks
• Preparation of competent cells (Using Calcium Chloride)
• Plasmid Extraction（Using Plasmid Mini Kit）
• Transformation (heat-shock method)
• Cleavage with two restriction enzymes
• Get Extraction
• Identification of DNA by Electrophoresis
• Ligation
• PCR of the colony
• Identification of DNA by Sequencing
• Others............
The forth layer assembly and assembled CHL1-3:
8/26 extract plasmid: 009-13,009-3. Enzyme digest 009-13,009- 3. Result shows that it is not cut completely .PCR 20 + 2 positive control.Transformation: 2 M-13 J-2 M-20 F, PSB, 2 K-16 MIV, CHL1-CHL3 (DH5 α and TrpR defects strains)
8/27 extract plasmid: 8H, 2M-14 H-2M. enzyme digest 009-3, 8H, 2 M-14 H-2 M. Ligate 009-2-3.Identification of the transformation of PCR 8/26 ligation results. Cut CHL1 religate CHL1-CHL3, CHL1-CHL4.Enzyme cut PCB, 23 L, RLS1.Transformation: CHL1-CHL3 (DH5 α and TrpR defects plant), CHL1-CHL4 (DH5 α and TrpR defects plant), 009-2-3, controls.
8/28 repeat 27th enzymes digest,ligated 009-2-3. Transformation: 18 M-RLS1, 18 O-RLS1, 2 G Ⅱ-RLS1, PSB-23 L, 009-2-3, CHL1-CHL3 (TrpR defects plant), CHL1-CHL4 (TrpR defects strains) controls. Reserve RLSF-2, PSB-1.....
Count Down
days
hours
minutes
seconds
Visitor
