Team:SDU-Denmark/Notebook
Notebook
Week 26
The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O. 5 µl diluted gblock and 95 µl ddH2O to make a 20X dilution. What we have done with the BioBrick K2018043 can be read in this protocol: Link to protocol (K2018043 - Cloning Basic part into iGEM standard plasmid psB1C3)
The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O. 5 µl diluted gblock and 95 µl ddH2O to make a 20X dilution. What we have done with the BioBrick K2018044 can be read in this protocol: Link to protocol (K2018044 - Cloning Basic part into iGEM standard plasmid psB1C3)
The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O. 5 µl diluted gblock and 95 µl ddH2O to make a 20X dilution. What we had done with the BioBrick K1763003 can be read in this protocol: Link to protocol (K1763003 - Cloning Basic part into iGEM standard plasmid psB1C3)
The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O. 5 µl diluted gblock and 95 µl ddH2O to make a 20X dilution. What we had done with the BioBrick K1763004 can be read in this protocol: Link to protocol (K1763004 - Cloning Basic part into iGEM standard plasmid psB1C3)
The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O. 5 µl diluted gblock and 95 µl ddH2O to make a 20X dilution. What we had done with the BioBrick K1763009 can be read in this protocol: Link to protocol (K1763009 - Cloning Basic part into iGEM standard plasmid psB1C3)
The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O. 5 µl diluted gblock and 95 µl ddH2O to make a 20X dilution. What we had done with the BioBrick K1763010 can be read in this protocol: Link to protocol (K1763010 - Cloning Basic part into iGEM standard plasmid psB1C3)
The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O. 5 µl diluted gblock and 95 µl ddH2O to make a 20X dilution. What we had done with the BioBrick K1763012 can be read in this protocol: Link to protocol (K1763012 - Cloning Basic part into iGEM standard plasmid psB1C3)
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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