Team:ZJU-China/labnote4.htm

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<h2>Week 4 (07.09-07.15): Make our own biobrick parts</h2>
<h2>Week 4 (07.09-07.15): Make our own biobrick parts</h2>
<h3>July 09</h3>
<h3>July 09</h3>
-
<p>Prepare liquid SOB medium for co-transformation.</p>
+
<p align="justify">Prepare liquid SOB medium for co-transformation.</p>
<h3>July 10</h3>
<h3>July 10</h3>
-
<p>Amplify glycerol stock bacteria FB (DH5&alpha;) on plates.</p>
+
<p align="justify">Amplify glycerol stock bacteria FB (DH5&alpha;) on plates.</p>
<h3>July 11</h3>
<h3>July 11</h3>
-
<p>1. Identify FA and FB with PCR.</p>
+
<p align="justify">1. Identify FA and FB with PCR.</p>
-
<p><img src="https://static.igem.org/mediawiki/igem.org/a/a2/Zju_notebook3.jpg"  width="500px"><p>
+
<p align="justify">
-
<p>2. Amplify FB-7-10-plate bacteria in liquid LB medium.</p>
+
<div class="floatC"><img src="https://static.igem.org/mediawiki/igem.org/a/a2/Zju_notebook3.jpg"  width="300px"><p align="justify"></div>
 +
<p align="justify">2. Amplify FB-7-10-plate bacteria in liquid LB medium.</p>
<h3>July 12</h3>
<h3>July 12</h3>
-
<p>1. Miniprep plasmid of pCJDFB from bacteria of July 06.</p>
+
<p align="justify">1. Miniprep plasmid of pCJDFB from bacteria of July 06.</p>
-
<p>Name: FB-7-11      Concentration: 15.4 ng/ul        A260/280: 1.88</p>
+
<p align="justify">Name: FB-7-11      Concentration: 15.4 ng/ul        A260/280: 1.88</p>
-
<p>2. Amplify FB-7-10-plate1, FB-7-10-plate2, FA-6.29-liq (DH5&alpha;), FA-lix and FB-lix bacteria for minipreparing plasmids later.</p>
+
<p align="justify">2. Amplify FB-7-10-plate1, FB-7-10-plate2, FA-6.29-liq (DH5&alpha;), FA-lix and FB-lix bacteria for minipreparing plasmids later.</p>
-
<p>3. Identify FB with PCR again.We found there were problems with primers used in those reactions, thus leading to the failure.</p>
+
<p align="justify">3. Identify FB with PCR again.We found there were problems with primers used in those reactions, thus leading to the failure.</p>
-
<p><img src="https://static.igem.org/mediawiki/igem.org/9/9f/Zju_notebook4.jpg"  width="500px"><p>
+
<p align="justify">
-
<p>4. Ligation: D0 and pSB1C3, purified at July 08.</p>
+
<div class="floatC"><img src="https://static.igem.org/mediawiki/igem.org/9/9f/Zju_notebook4.jpg"  width="300px"><p align="justify"></div>
 +
<p align="justify">4. Ligation: D0 and pSB1C3, purified at July 08.</p>
<h3>July 13</h3>
<h3>July 13</h3>
-
<p>1. Transformation:</p>
+
<p align="justify">1. Transformation:</p>
-
<p>No.name  time of heat shock/s</p>
+
<p align="justify">No.name  time of heat shock/s</p>
-
<p>1  D0-FA-FB 90</p>
+
<p align="justify">1  D0-FA-FB 90</p>
-
<p>2  FA-FB 90</p>
+
<p align="justify">2  FA-FB 90</p>
-
<p>3  D0-FA-FB 30</p>
+
<p align="justify">3  D0-FA-FB 30</p>
-
<p>4  FA-FB 30</p>
+
<p align="justify">4  FA-FB 30</p>
-
<p>5  ligation product 90</p>
+
<p align="justify">5  ligation product 90</p>
-
<p>6  D0 90</p>
+
<p align="justify">6  D0 90</p>
-
<p>2. Miniprep plasmid of FA and FB from bacteria of July 12.</p>
+
<p align="justify">2. Miniprep plasmid of FA and FB from bacteria of July 12.</p>
-
<p>Name: FA-7-13-1-1      Concentration: 15.8 ng/ul        A260/280: 1.89</p>
+
<p align="justify">Name: FA-7-13-1-1      Concentration: 15.8 ng/ul        A260/280: 1.89</p>
-
<p>Name: FA-7-13-1-2      Concentration: 10.8 ng/ul        A260/280: 2.06</p>
+
<p align="justify">Name: FA-7-13-1-2      Concentration: 10.8 ng/ul        A260/280: 2.06</p>
-
<p>Name: FA-7-13-1-3      Concentration: 13.6 ng/ul        A260/280: 2.30</p>
+
<p align="justify">Name: FA-7-13-1-3      Concentration: 13.6 ng/ul        A260/280: 2.30</p>
-
<p>Name: FA-7-13-1-4      Concentration: 12.3 ng/ul        A260/280: 2.13</p>
+
<p align="justify">Name: FA-7-13-1-4      Concentration: 12.3 ng/ul        A260/280: 2.13</p>
-
<p>Name: FA-7-13-2-1      Concentration: 9.4 ng/ul          A260/280: 2.15</p>
+
<p align="justify">Name: FA-7-13-2-1      Concentration: 9.4 ng/ul          A260/280: 2.15</p>
-
<p>Name: FA-7-13-2-2      Concentration: 12.4 ng/ul        A260/280: 1.94</p>
+
<p align="justify">Name: FA-7-13-2-2      Concentration: 12.4 ng/ul        A260/280: 1.94</p>
-
<p>Name: FA-7-13-2-3      Concentration: 20.1 ng/ul        A260/280: 1.94</p>
+
<p align="justify">Name: FA-7-13-2-3      Concentration: 20.1 ng/ul        A260/280: 1.94</p>
-
<p>Name: FA-7-13-3-1      Concentration: 11.1 ng/ul        A260/280: 2.23</p>
+
<p align="justify">Name: FA-7-13-3-1      Concentration: 11.1 ng/ul        A260/280: 2.23</p>
-
<p>Name: FA-7-13-lix      Concentration: 13.3 ng/ul        A260/280: 2.08</p>
+
<p align="justify">Name: FA-7-13-lix      Concentration: 13.3 ng/ul        A260/280: 2.08</p>
-
<p>Name: FB-7-13-yy      Concentration: 26.4 ng/ul        A260/280: 1.84</p>
+
<p align="justify">Name: FB-7-13-yy      Concentration: 26.4 ng/ul        A260/280: 1.84</p>
-
<p><img src="https://static.igem.org/mediawiki/igem.org/8/85/Zju_notebook5.jpg"  width="80%"><p>
+
<p align="justify">
 +
<div class="floatC"><img src="https://static.igem.org/mediawiki/igem.org/8/85/Zju_notebook5.jpg"  width="300px"><p align="justify"></div>
<h3>July 14</h3>
<h3>July 14</h3>
-
<p>1. Co-transformation: </p>
+
<p align="justify">1. Co-transformation: </p>
-
<p>No.name  time of heat shock/s</p>
+
<p align="justify">No.name  time of heat shock/s</p>
-
<p>1  D0-FA-FB  90</p>
+
<p align="justify">1  D0-FA-FB  90</p>
-
<p>2  D0-FA-FB  30</p>
+
<p align="justify">2  D0-FA-FB  30</p>
-
<p>Use SOC liquid medium.</p>
+
<p align="justify">Use SOC liquid medium.</p>
-
<p>2. Miniprep plasmid of FA and FB from bacteria of July 12.</p>
+
<p align="justify">2. Miniprep plasmid of FA and FB from bacteria of July 12.</p>
-
<p>Name: FB-7-14-1      Concentration: 42.9 ng/ul        A260/280: 0.67</p>
+
<p align="justify">Name: FB-7-14-1      Concentration: 42.9 ng/ul        A260/280: 0.67</p>
-
<p>Name: FB-7-14-2      Concentration: 21.0 ng/ul        A260/280: 2.69</p>
+
<p align="justify">Name: FB-7-14-2      Concentration: 21.0 ng/ul        A260/280: 2.69</p>
-
<p>Name: FB-7-14-3      Concentration: 44.7 ng/ul        A260/280: 0.59</p>
+
<p align="justify">Name: FB-7-14-3      Concentration: 44.7 ng/ul        A260/280: 0.59</p>
-
<p>Name: FB-7-14-4      Concentration: 14.4 ng/ul        A260/280: 2.09</p>
+
<p align="justify">Name: FB-7-14-4      Concentration: 14.4 ng/ul        A260/280: 2.09</p>
-
<p>Name: FB-7-14-5      Concentration: 33.1 ng/ul        A260/280: 0.68</p>
+
<p align="justify">Name: FB-7-14-5      Concentration: 33.1 ng/ul        A260/280: 0.68</p>
-
<p>Name: FB-7-14-6      Concentration: 26.1 ng/ul        A260/280: 1.64</p>
+
<p align="justify">Name: FB-7-14-6      Concentration: 26.1 ng/ul        A260/280: 1.64</p>
-
<p><img src="https://static.igem.org/mediawiki/igem.org/3/3e/Zju_notebook6.jpg"  width="500px"><p>
+
<p align="justify"><div class="floatC"><img src="https://static.igem.org/mediawiki/igem.org/3/3e/Zju_notebook6.jpg"  width="300px"><p align="justify"></div>
-
<p>3. Amplify glycerol stock bacteria co-transformation (BL21*(DE3)) on 3 resistance plates, glycerol stock bacteria D0 on Amp plate.</p>
+
<p align="justify">3. Amplify glycerol stock bacteria co-transformation (BL21*(DE3)) on 3 resistance plates, glycerol stock bacteria D0 on Amp plate.</p>
-
<p>4. Amplify FB-7-12-2, FB-7-12-4, FA-7-12-2-2 and FA-7-12-2-3 in LB liquid medium, named FB-7-14-2, FB-7-14-4, FA-7-14-2-2 and FA-7-14-2-3.</p>
+
<p align="justify">4. Amplify FB-7-12-2, FB-7-12-4, FA-7-12-2-2 and FA-7-12-2-3 in LB liquid medium, named FB-7-14-2, FB-7-14-4, FA-7-14-2-2 and FA-7-14-2-3.</p>
-
<p>5. Digest FA-7-13-2-2, FA-7-13-2-3, FB-7-14-2 and FB-7-14-4 with Pst1.</p>
+
<p align="justify">5. Digest FA-7-13-2-2, FA-7-13-2-3, FB-7-14-2 and FB-7-14-4 with Pst1.</p>
-
<p><img src="https://static.igem.org/mediawiki/igem.org/d/de/Zju_notebook7.jpg"  width="500px"><p>
+
<p align="justify"><div class="floatC"><img src="https://static.igem.org/mediawiki/igem.org/d/de/Zju_notebook7.jpg"  width="300px"><p align="justify"></div>
-
<p>Maybe the concentration is too low that we can't recognize it.</p>
+
<p align="justify">Maybe the concentration is too low that we can't recognize it.</p>
<h3>July 15</h3>
<h3>July 15</h3>
-
<p>1. Transformation:</p>
+
<p align="justify">1. Transformation:</p>
-
<p>No.  name  time of heat shock/s  Strain</p>
+
<p align="justify">No.  name  time of heat shock/s  Strain</p>
-
<p>1  D0-FA-FB  90  DH10&beta;</p>
+
<p align="justify">1  D0-FA-FB  90  DH10&beta;</p>
-
<p>2  FA-FB  90  DH10&beta;</p>
+
<p align="justify">2  FA-FB  90  DH10&beta;</p>
-
<p>3  D0-FA-FB  30  BL21*(DE3)</p>
+
<p align="justify">3  D0-FA-FB  30  BL21*(DE3)</p>
-
<p>4  FA-FB  30  BL21*(DE3)</p>
+
<p align="justify">4  FA-FB  30  BL21*(DE3)</p>
-
<p>5  D0-pSB1C3  90  BL21*(DE3)</p>
+
<p align="justify">5  D0-pSB1C3  90  BL21*(DE3)</p>
-
<p>6  D0  90  BL21*(DE3)</p>
+
<p align="justify">6  D0  90  BL21*(DE3)</p>
-
<p>2. Amplify bacteria in LB liquid medium:</p>
+
<p align="justify">2. Amplify bacteria in LB liquid medium:</p>
-
<p>No.  Name  From  quantity</p>
+
<p align="justify">No.  Name  From  quantity</p>
-
<p>1  Co-7-15  co-transformation (BL21*(DE3))  6</p>
+
<p align="justify">1  Co-7-15  co-transformation (BL21*(DE3))  6</p>
-
<p>2  D0-7-15  D0-7-14  1</p>
+
<p align="justify">2  D0-7-15  D0-7-14  1</p>
-
<p>3  Co3-7-15-1-3-1  Co3-7-14-1  1</p>
+
<p align="justify">3  Co3-7-15-1-3-1  Co3-7-14-1  1</p>
-
<p>4  Co3-7-15-1-3-2  Co3-7-14-2  1</p>
+
<p align="justify">4  Co3-7-15-1-3-2  Co3-7-14-2  1</p>
</body></html>
</body></html>

Latest revision as of 17:13, 26 October 2012

HOME

Week 4 (07.09-07.15): Make our own biobrick parts

July 09

Prepare liquid SOB medium for co-transformation.

July 10

Amplify glycerol stock bacteria FB (DH5α) on plates.

July 11

1. Identify FA and FB with PCR.

2. Amplify FB-7-10-plate bacteria in liquid LB medium.

July 12

1. Miniprep plasmid of pCJDFB from bacteria of July 06.

Name: FB-7-11 Concentration: 15.4 ng/ul A260/280: 1.88

2. Amplify FB-7-10-plate1, FB-7-10-plate2, FA-6.29-liq (DH5α), FA-lix and FB-lix bacteria for minipreparing plasmids later.

3. Identify FB with PCR again.We found there were problems with primers used in those reactions, thus leading to the failure.

4. Ligation: D0 and pSB1C3, purified at July 08.

July 13

1. Transformation:

No.name time of heat shock/s

1 D0-FA-FB 90

2 FA-FB 90

3 D0-FA-FB 30

4 FA-FB 30

5 ligation product 90

6 D0 90

2. Miniprep plasmid of FA and FB from bacteria of July 12.

Name: FA-7-13-1-1 Concentration: 15.8 ng/ul A260/280: 1.89

Name: FA-7-13-1-2 Concentration: 10.8 ng/ul A260/280: 2.06

Name: FA-7-13-1-3 Concentration: 13.6 ng/ul A260/280: 2.30

Name: FA-7-13-1-4 Concentration: 12.3 ng/ul A260/280: 2.13

Name: FA-7-13-2-1 Concentration: 9.4 ng/ul A260/280: 2.15

Name: FA-7-13-2-2 Concentration: 12.4 ng/ul A260/280: 1.94

Name: FA-7-13-2-3 Concentration: 20.1 ng/ul A260/280: 1.94

Name: FA-7-13-3-1 Concentration: 11.1 ng/ul A260/280: 2.23

Name: FA-7-13-lix Concentration: 13.3 ng/ul A260/280: 2.08

Name: FB-7-13-yy Concentration: 26.4 ng/ul A260/280: 1.84

July 14

1. Co-transformation:

No.name time of heat shock/s

1 D0-FA-FB 90

2 D0-FA-FB 30

Use SOC liquid medium.

2. Miniprep plasmid of FA and FB from bacteria of July 12.

Name: FB-7-14-1 Concentration: 42.9 ng/ul A260/280: 0.67

Name: FB-7-14-2 Concentration: 21.0 ng/ul A260/280: 2.69

Name: FB-7-14-3 Concentration: 44.7 ng/ul A260/280: 0.59

Name: FB-7-14-4 Concentration: 14.4 ng/ul A260/280: 2.09

Name: FB-7-14-5 Concentration: 33.1 ng/ul A260/280: 0.68

Name: FB-7-14-6 Concentration: 26.1 ng/ul A260/280: 1.64

3. Amplify glycerol stock bacteria co-transformation (BL21*(DE3)) on 3 resistance plates, glycerol stock bacteria D0 on Amp plate.

4. Amplify FB-7-12-2, FB-7-12-4, FA-7-12-2-2 and FA-7-12-2-3 in LB liquid medium, named FB-7-14-2, FB-7-14-4, FA-7-14-2-2 and FA-7-14-2-3.

5. Digest FA-7-13-2-2, FA-7-13-2-3, FB-7-14-2 and FB-7-14-4 with Pst1.

Maybe the concentration is too low that we can't recognize it.

July 15

1. Transformation:

No. name time of heat shock/s Strain

1 D0-FA-FB 90 DH10β

2 FA-FB 90 DH10β

3 D0-FA-FB 30 BL21*(DE3)

4 FA-FB 30 BL21*(DE3)

5 D0-pSB1C3 90 BL21*(DE3)

6 D0 90 BL21*(DE3)

2. Amplify bacteria in LB liquid medium:

No. Name From quantity

1 Co-7-15 co-transformation (BL21*(DE3)) 6

2 D0-7-15 D0-7-14 1

3 Co3-7-15-1-3-1 Co3-7-14-1 1

4 Co3-7-15-1-3-2 Co3-7-14-2 1

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