Difference between revisions of "Team:CAU China/Notebook"

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<p>Digest the mixture with restriction enzymes: XbaⅠ-HF and SacⅠ</p>
 
<p>Digest the mixture with restriction enzymes: XbaⅠ-HF and SacⅠ</p>
 
<img src="https://static.igem.org/mediawiki/2015/2/2a/CAU_notebook_2.JPG" width="400px">
 
<img src="https://static.igem.org/mediawiki/2015/2/2a/CAU_notebook_2.JPG" width="400px">
 +
<p>After four-hour digest, the product was purified with the silicagel column</p>
 +
<p>Linkage</p>
 +
<img src="https://static.igem.org/mediawiki/2015/8/87/CAU_notebook_3.JPG" width="400px">
 +
<p>Link overnight</p>
 +
<p>The product was named as pHSE-A</p>
 +
<p>Saturday 4/4</p>
 +
<p>Transform the Escherichia Coli (strain EP1300) with pHSE-A</p>
 +
<p>Mix EP1300 and pHSE-A, 0℃ice bath 30min</p>
 +
<p>42℃ water bath 90s</p>
 +
<p>0℃ice bath 1min</p>
 +
<p>37℃recover 1h</p>
 +
<p>Sunday 5/4</p>
 +
<p>There were many colonies growing up on the plate</p>
 +
<p>Chose ten of them and scribed on another LB plate which contained kanamycin (K100)</p>
  
 +
<p>Monday 6/4</p>
 +
<p>Three bacterial colonies from the agar plate from yesterday were transferred to 3 bottle of fluid LB medium to amplify bacteria</p>
 +
<p>Extract the pHSE-A plasmid from the bacteria</p>
 +
<p>The plasmid was identified by enzyme digestion——XbaⅠ&SacⅠ, NcoⅠ&StuⅠ</p>
 +
<img src="https://static.igem.org/mediawiki/2015/b/b2/CAU_notebook_4.JPG" width="400px">
 +
<p>The clone was right</p>
 +
 +
<p>Tuesday 7/4</p>
 +
<p>Digest the pHSE-A and pUC57-BT/TB/BAG/GAB respectively with XbaⅠ-HF and StuⅠ</p>
 +
<img src="https://static.igem.org/mediawiki/2015/4/40/CAU_notebook_5.JPG" width="400px">
 +
<p>After four-hour digest, the products were then analysed on gel-electrophoresis and purified with the silicagel column</p>
 +
<img src="https://static.igem.org/mediawiki/2015/e/ee/CAU_notebook_6.JPG" width="400px">
  
 
</html>
 
</html>

Revision as of 04:50, 6 September 2015

Notebook

Lab Journal

"Do not, for one repulse, forgot the purpose that you resolved to effort." - William Shakespeare

Wednesday 1/4

We started our work in the wet lab with a one-day lab safety course. We learned basic lab safety knowledge, such as how to put out a fire in the lab, how to deal with the poisonous chemical and the dangerous bacteria.

Thursday 2/4

We started to get familiar with our lab. We learned about where we can find everything that is needed for working in the lab. In addition, we had a lab meeting about the project goals and the way how to get there.

Friday 3/4

PCR reaction on pER41G-A

Gel purification was performed on the PCR product

The product was named as Ap-PCR(P)

Mix the Ap-PCR(P) and pER41G-A

Digest the mixture with restriction enzymes: XbaⅠ-HF and SacⅠ

After four-hour digest, the product was purified with the silicagel column

Linkage

Link overnight

The product was named as pHSE-A

Saturday 4/4

Transform the Escherichia Coli (strain EP1300) with pHSE-A

Mix EP1300 and pHSE-A, 0℃ice bath 30min

42℃ water bath 90s

0℃ice bath 1min

37℃recover 1h

Sunday 5/4

There were many colonies growing up on the plate

Chose ten of them and scribed on another LB plate which contained kanamycin (K100)

Monday 6/4

Three bacterial colonies from the agar plate from yesterday were transferred to 3 bottle of fluid LB medium to amplify bacteria

Extract the pHSE-A plasmid from the bacteria

The plasmid was identified by enzyme digestion——XbaⅠ&SacⅠ, NcoⅠ&StuⅠ

The clone was right

Tuesday 7/4

Digest the pHSE-A and pUC57-BT/TB/BAG/GAB respectively with XbaⅠ-HF and StuⅠ

After four-hour digest, the products were then analysed on gel-electrophoresis and purified with the silicagel column