2405-pEC-Miraculin-Term
Cloning of 2412 pEC2-AaeMutS
[edit]Purpose: Cloning of 2412 pEC2-AaeMutS
Date: 2015-3-5
Notebook:
Page:
Clone ID: 2412
Experimenters: Paul & Xu Xin
Results
[edit]cloning
[edit]1.Use plant as template to extract genomic DNA.
2. PCR use the genomic DNA as template.
The 1st: the DNA extacted by Paul; the 2nd PCR: The DNA extacted by Xin
3. Gel purified the PCR#2,
4. Digestion, clean up digestion (with NcoI/KpnI), ligation. transformaiton.
5. select 10 colonies from transformation plate to PCR screening.
6. Select colonies 1,2,7,8,9 and 10 to innoculate.
7. extact plasmid from the step 6.
8. Digestion the plasmids with NcoI/KpnI
9. From the gel, no insection in these 6 plasmid.
10. As Paul curious what is the product,PCR the these 6 selected plasmids. Also PCR the genmoic DNA as step 1.
Send plasmid NO. 7 to sequence.
Also the genomic DNA PCR was purified, but elution in 40ul (instead of 20) and casue to low concentration ( be careful for the next step), maybe need to rewash and elution in a low concentration.
Date: