3257 pCMV5-anti-mkappa-rFc-His
Cloning of 3257 pCMV5-anti-mkappa-rFc-His
[edit]Purpose: Cloning of 3257 pCMV5-anti-mkappa-rFc-His
Date: 08, 15, 2018
Notebook:
Page:
Clone ID: 3257
Experimenters: yijia
Results
[edit]Cloning
[edit]gene synthesis of 3257-F1
template:
1st PCR of 3257-F2
PCR from 1413 pCDNA3.4-Rabbit-IgG Heavy Chain
3257-F2 CACTGGTCACAGTGTCTAGCGCTAGCGGTACCAGCAAGCCCACGTGCCCAC
3257-Not1-R3 TTGATTGCGGCCGCTCAGCAGTGATGGTGATGATGATGTTTACCCGGAGAGCGGGAGATG
2nd PCR of 3257
combine both gene synthesis fragments and rabbit Fc fragment
Purify with column PCR again using
3257-1 TGTCACGAATTCACAGGTGCAGCTGGTG
3257-Not1-R3 TTGATTGCGGCCGCTCAGCAGTGATGGTGATGATGATGTTTACCCGGAGAGCGGGAGATG
1160bp
Vector digestion
Vector is 3075
Colony screen
[edit]3257 PCR Screening
Miniprep of 3257
| # | Lane | Enzymes | Length | Buffer | Note |
| 3257 | 1-4(#1-4) | EcoR1+Not1 | 1160bp | Fastdigest | #2 to do sequence |
| vector control | 3075 | EcoR1+Not1 | 681bp | Fastddigest |
10/16/2018: in order to digest with Nhe1, plasmid has to be isolated from INV110.
Result intepretation/Summary
[edit]Conclusion
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