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 BioBrick was ordered from IDT in a plasmid pUCIDT-AMP and after receiving it, it was resuspended to a final concentration of 0.1 µg/µl. After the resuspension, the plasmid was transformed into Top10 E. coli by following SOP0023_v01, with the following remarks: 2.5 µl plasmid was used instead of the normal 1 µl. What we had done with the BioBrick K2018010 can be read in this protocol: Link to protocol (K2018010 - Cloning composite part into iGEM standard plasmid) and (IMPACT Purification of K2018010)
The BioBrick 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.
What we had done with the BioBrick K2018011 can be read in this protocol: Link to protocol (K2018011 - Cloning composite part into iGEM standard plasmid), (Cloning Thuricin with DA silk-overhangs into pSB1C3), (Cloning Thuricin with DE silk-overhangs into pSB1C3), (Cloning Thuricin with EA silk-overhangs into pSB1C3), (Cloning Thuricin with ED silk-overhangs into pSB1C3) and (IMPACT Purification of K2018011)
The BioBrick was ordered from IDT in a plasmid pUCIDT-AMP and after receiving it, it was resuspended to a final concentration of 0.1 µg/µl. After the resuspension, the plasmid was transformed into Top10 E. coli by following SOP0023_v01, with the following remarks: 2.5 µl plasmid was used instead of the normal 1 µl. What we had done with the BioBrick K2018014 can be read in this protocol: Link to protocol (K2018014 - Cloning composite part into iGEM standard plasmid) and (IMPACT Purification of K2018014)
The BioBrick was ordered from IDT in a plasmid pUCIDT-AMP and after receiving it, it was resuspended to a final concentration of 0.1 µg/µl. After the resuspension, the plasmid was transformed into Top10 E. coli by following SOP0023_v01, with the following remarks: 2.5 µl plasmid was used instead of the normal 1 µl. What we had done with the BioBrick K2018019 can be read in this protocol: Link to protocol (K2018019 - Cloning composite part into iGEM standard plasmid) and (IMPACT Purification of K2018019)
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