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Team:ZJU-China/labnote7.htm
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<p>Name: K738000-7-30-10 Concentration: 43.6 ng/ul A260/280: 1.98</p> | <p>Name: K738000-7-30-10 Concentration: 43.6 ng/ul A260/280: 1.98</p> | ||
<p>PCR of K738000-7-30</p> | <p>PCR of K738000-7-30</p> | ||
| - | <p><img src="https://static.igem.org/mediawiki/igem.org/d/df/Zju_notebook15.jpg" width=" | + | <p><img src="https://static.igem.org/mediawiki/igem.org/d/df/Zju_notebook15.jpg" width="500px"><p> |
<p>Unfortunately, there was a slight electrophoretic band in control. But the sequencing result came later verified that we've got the correct ligation product.</p> | <p>Unfortunately, there was a slight electrophoretic band in control. But the sequencing result came later verified that we've got the correct ligation product.</p> | ||
<p>2. Pick up single colonies from plate of FB-7-26-2 and amplify it in LB liquid medium.</p> | <p>2. Pick up single colonies from plate of FB-7-26-2 and amplify it in LB liquid medium.</p> | ||
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<p>Name: K738000-7-30-11 Concentration: 46.0 ng/ul A260/280: 1.97</p> | <p>Name: K738000-7-30-11 Concentration: 46.0 ng/ul A260/280: 1.97</p> | ||
<p>Name: FA-7-28-1 Concentration: 33.0 ng/ul A260/280: 2.04</p> | <p>Name: FA-7-28-1 Concentration: 33.0 ng/ul A260/280: 2.04</p> | ||
| - | <p><img src="https://static.igem.org/mediawiki/igem.org/e/e6/Zju_notebook16.jpg" width=" | + | <p><img src="https://static.igem.org/mediawiki/igem.org/e/e6/Zju_notebook16.jpg" width="500px"><p> |
<h3>August 01</h3> | <h3>August 01</h3> | ||
<p>Double digest K738000-7-23-2 and K738000-7-23-3 with EcoR1 and Pst1 and got linear backbone through DNA gel extraction.</p> | <p>Double digest K738000-7-23-2 and K738000-7-23-3 with EcoR1 and Pst1 and got linear backbone through DNA gel extraction.</p> | ||
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<p>Name: FA-8-4 Concentration: 19.6 ng/ul A260/280: 1.97</p> | <p>Name: FA-8-4 Concentration: 19.6 ng/ul A260/280: 1.97</p> | ||
<p>Name: FB-8-4 Concentration: 35.9 ng/ul A260/280: 1.98</p> | <p>Name: FB-8-4 Concentration: 35.9 ng/ul A260/280: 1.98</p> | ||
| - | <p><img src="https://static.igem.org/mediawiki/igem.org/c/cc/Zju_notebook17.jpg" width=" | + | <p><img src="https://static.igem.org/mediawiki/igem.org/c/cc/Zju_notebook17.jpg" width="500px"><p> |
<p>Unfortunately, the PCR results were so different from that of original plasmids. We couldn't figure out the reason.</p> | <p>Unfortunately, the PCR results were so different from that of original plasmids. We couldn't figure out the reason.</p> | ||
<p>2. Amplify Venus-8-2 in LB liquid medium, named Venus-8-4.</p> | <p>2. Amplify Venus-8-2 in LB liquid medium, named Venus-8-4.</p> | ||
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<p>1. Pick up single colonies from plate of FAM-8-4 and FBP-8-4 and amplify it in LB liquid medium and LB plates.</p> | <p>1. Pick up single colonies from plate of FAM-8-4 and FBP-8-4 and amplify it in LB liquid medium and LB plates.</p> | ||
<p>2. Colony PCR of FAM-8-4-2 and FBP-8-4-2</p> | <p>2. Colony PCR of FAM-8-4-2 and FBP-8-4-2</p> | ||
| - | <p><img src="https://static.igem.org/mediawiki/igem.org/c/c8/Zju_notebook18.jpg" width=" | + | <p><img src="https://static.igem.org/mediawiki/igem.org/c/c8/Zju_notebook18.jpg" width="500px"><p> |
<p>3. Double digest BB-8-4-1 with (1) EcoR1 and Pst1, (2) Xba1 and Spe1, get linear backbone through PCR cleanup kit.</p> | <p>3. Double digest BB-8-4-1 with (1) EcoR1 and Pst1, (2) Xba1 and Spe1, get linear backbone through PCR cleanup kit.</p> | ||
<p>4. Miniprep plasmid of Venus-8-4.</p> | <p>4. Miniprep plasmid of Venus-8-4.</p> | ||
Revision as of 13:13, 25 September 2012
Week 7 (07.30-08.05): Test part BBa_K537009 and make our own parts
July 30
1. Miniprep plasmid of K738000-7-29.
Name: K738000-7-30-1 Concentration: 60.0 ng/ul A260/280: 1.86
Name: K738000-7-30-2 Concentration: 47.1 ng/ul A260/280: 1.95
Name: K738000-7-30-3 Concentration: 90.1 ng/ul A260/280: 1.95
Name: K738000-7-30-10 Concentration: 43.6 ng/ul A260/280: 1.98
PCR of K738000-7-30
Unfortunately, there was a slight electrophoretic band in control. But the sequencing result came later verified that we've got the correct ligation product.
2. Pick up single colonies from plate of FB-7-26-2 and amplify it in LB liquid medium.
July 31
1. Amplify backbone pSB1C3:
Double digest K738000-7-23-3 with EcoR1 and Pst1 and got linear backbone through DNA gel extraction.
2. Miniprep plasmid of K738000-7-29 and FA-7-28-1.
Name: K738000-7-30-4 Concentration: 83.9 ng/ul A260/280: 1.94
Name: K738000-7-30-5 Concentration: 50.1 ng/ul A260/280: 2.00
Name: K738000-7-30-11 Concentration: 46.0 ng/ul A260/280: 1.97
Name: FA-7-28-1 Concentration: 33.0 ng/ul A260/280: 2.04
August 01
Double digest K738000-7-23-2 and K738000-7-23-3 with EcoR1 and Pst1 and got linear backbone through DNA gel extraction.
August 02
1. Transform part K537009 into DH5α to acquire experience, named venus-8-2.
2. Transform original pCJDFA and pCJDFB into DH5α, named FA-8-2 and FB-8-2.
3. Transform part K398326 into DH10β to amplify backbone pSB1C3 easier, BB-8-2.
August 03
1. Amplify BB-8-2, FA-8-2 and FB -8-2 for minipreparing plasmids later, named BB-8-3, FA-8-3 and FB-8-3.
August 04
1. Miniprep plasmid of BB-8-3 FA-8-3 and FB-8-3.
Name: BB-8-4 Concentration: 76.2 ng/ul A260/280: 1.91
Name: BB-8-4 Concentration: 40.3 ng/ul A260/280: 1.90
Name: FA-8-4 Concentration: 19.6 ng/ul A260/280: 1.97
Name: FB-8-4 Concentration: 35.9 ng/ul A260/280: 1.98
Unfortunately, the PCR results were so different from that of original plasmids. We couldn't figure out the reason.
2. Amplify Venus-8-2 in LB liquid medium, named Venus-8-4.
3. Transform FAM-8-4-1, FAM-8-4-1, FBP-8-4-1 and FBP-8-4-1 into DH5α.
August 05
1. Pick up single colonies from plate of FAM-8-4 and FBP-8-4 and amplify it in LB liquid medium and LB plates.
2. Colony PCR of FAM-8-4-2 and FBP-8-4-2
3. Double digest BB-8-4-1 with (1) EcoR1 and Pst1, (2) Xba1 and Spe1, get linear backbone through PCR cleanup kit.
4. Miniprep plasmid of Venus-8-4.
Name: Venus-8-5-1 Concentration: 168.1 ng/ul A260/280: 1.91
Name: Venus-8-5-1 Concentration: 83.4 ng/ul A260/280: 1.93
5. Pick up single colonies from plate of FAM-8-4-1, FAM-8-4-2, FBP-8-4-1 and FBP-8-4-2 and amplify it in LB liquid medium and LB plates. Miniprep plasmid of them.
Name: FAM-8-5-1 Concentration: 69.9 ng/ul A260/280: 1.95
Name: FAM-8-5-2 Concentration: 57.3 ng/ul A260/280: 1.95
Name: FAM-8-5-3 Concentration: 86.8 ng/ul A260/280: 2.06
Name: FBP-8-5-2 Concentration: 21.0 ng/ul A260/280: 2.14
Name: FBP-8-5-3 Concentration: 15.5 ng/ul A260/280: 2.14
Name: FBP-8-5-5 Concentration: 15.6 ng/ul A260/280: 2.07
6. Co-transform FA-8-5-3, FB-8-5-2 and D0-4 into BL21*(DE3). Co-transform FA-8-5-3 and FB-8-5-2 into BL21*(DE3).
