Difference between revisions of "Team:CAU China/Notebook"
| Line 46: | Line 46: | ||
<p>After four-hour digest, the products were then analysed on gel-electrophoresis and purified with the silicagel column</p> | <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"> | <img src="https://static.igem.org/mediawiki/2015/e/ee/CAU_notebook_6.JPG" width="400px"> | ||
| + | <p>The products were named as pHSE-A-XS and BT/TB/BAG/GAB-XS</p> | ||
| + | <p>Linkage</p> | ||
| + | <img src="https://static.igem.org/mediawiki/2015/b/bd/CAU_notebook_7.JPG" width="400px"> | ||
| + | <p>Link overnight</p> | ||
| + | <p>The products were named as pHSE-BT/TB/BAG/GAB</p> | ||
| + | |||
| + | <p>Wednesday 8/4</p> | ||
| + | <p>Transform the Escherichia Coli (strain EP1300) with pHSE-BT/TB/BAG/GAB</p> | ||
| + | <p>The transformed strain was plated out on the solid LB medium which contained kanamycin (K100)</p> | ||
| + | <p>Thursday 9/4</p> | ||
| + | <p>Choose about 10 of the grown-up colonies respectively and scribed on another LB plate which contained ampicillin (A100) for invert selection</p> | ||
| + | |||
| + | <p>Friday 10/4</p> | ||
| + | <p>Choose the colonies which can grow on the kanamycin plate but can’t grow on the ampicillin plate</p> | ||
| + | <p>Colony PCR analysis</p> | ||
| + | <img src="https://static.igem.org/mediawiki/2015/b/be/CAU_notebook_8.JPG" width="400px"> | ||
| + | <p>Two bacterial colonies from each agar plate were transferred to 8 bottle of fluid LB medium to amplify bacteria.</p> | ||
| + | <p>Extract the plasmid from the bacteria</p> | ||
| + | <p>The plasmid was identified by enzyme digestion——XbaⅠ-HF and StuⅠ</p> | ||
| + | <img src="https://static.igem.org/mediawiki/2015/a/a2/CAU_notebook_9.JPG" width="400px"> | ||
</html> | </html> | ||
Revision as of 05:06, 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
The products were named as pHSE-A-XS and BT/TB/BAG/GAB-XS
Linkage
Link overnight
The products were named as pHSE-BT/TB/BAG/GAB
Wednesday 8/4
Transform the Escherichia Coli (strain EP1300) with pHSE-BT/TB/BAG/GAB
The transformed strain was plated out on the solid LB medium which contained kanamycin (K100)
Thursday 9/4
Choose about 10 of the grown-up colonies respectively and scribed on another LB plate which contained ampicillin (A100) for invert selection
Friday 10/4
Choose the colonies which can grow on the kanamycin plate but can’t grow on the ampicillin plate
Colony PCR analysis
Two bacterial colonies from each agar plate were transferred to 8 bottle of fluid LB medium to amplify bacteria.
Extract the plasmid from the bacteria
The plasmid was identified by enzyme digestion——XbaⅠ-HF and StuⅠ
