Notebook
Week 28
The brick containing the gene K2018011 was digested with PstI and XbaI overnight by using SOP0017_v01.
After digesting, the sample was ligated by following SOP0015_v01 with the standard iGEM plasmid, pSB1C3, overnight, and transformed into E.coli Top10.
cPCR program used:
Segment | Step | Temperature | Duration |
---|---|---|---|
1 | Initial denaturation | 98 °C | 2 min |
2 | 34 cycles | 98 °C | 10 sec |
72 °C | 15 sec | ||
72 °C | >15 sec | ||
3 | Final extension | 72 °C | 5 min |
4 | Keep the sample cold | 12 °C | HOLD |
After a colony PCR, SOP0021_v01, was run and verified on a gel, the following result was given: Our part is 259 bp and primer overhangs are 150 X 2 bp, which is, in total, 559 bp: A gel electrophoresis from cPCR showed a band around 600 bp, so we assumed that we had the correct part inserted into the iGEM standard plasmid.
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