Difference between revisions of "Team:SDU-Denmark/Notebook"

Revision as of 21:22, 14 October 2016

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

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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.