13 July
Designed Primers for Taq into backbone
Researched about using an Asp/Pro between the peptide and the Taq
Began looking at ligation independent cloning for insertion of peptide
23 July
Checked thermostability of the mutations that will be done to Taq using Expasy
Sent in sequence of taq with mutations at 485, 515 and 540 to iTasser
3 August
Had iGEM team meeting
-Discussed ANZAAS Proposal
-Spoke about possibilities of using Geneart
-Discussed possible T-shirt designs
5 August
Meet up with Damian, discussed different strategies to use in modelling, began using YASARA to do our Modelling
10 August
Had iGEM team meeting
-Took Team Photo
11 August
PCR of Taq
Taq with a DNA concentration of 40ng/µL was diluted to 1µM by addition of 24µL of MilliQ water to 1µL of the Taq plasmid. Primers were diluted to 100µM by adding 354µL to the forward primer with a concentration of 35.4 nmol and 405µL to the reverser primer with a concentration of 40.5nmol. The primers were further diluted to 0.2µM by adding 10µL of the primer to 490µL of milliQ water. The PCR reaction was then made by adding 25µL of Master Mix, 2.5µL of each primer, 5µL of Rnase free water and 15µL of 0.2µM Taq Plasmid.
The PCR
98º 60 Sec 1 cycle
98º 15 Sec then 72º 90 Sec 25 cycle
72º 5 min 1 cycle
16 August
Collaborated with the Sheffield team by participating in an over the phone survery about Human Practices
17 August
had team meeting
- discussed how we can get more sponsors
- talked about placing our order to gene art
- began filling out the form to send Pennstates survey to the ethics committee
- discussed results for modelling
- decided to fix optimise our previous PCR
18 August
called few companies seeking sponsorship
enquired for booking flights and accomadation
Working on comparing TAQ wild structure to the TAQ Mutated structure to see that there are no major differences, which will mean that TAQ is inactive but will retain it's thermostability at high temperatures.
Wrote an email to ABC TV, wanting to know if they will do piece on our project for there The New Inventors Program.
Waiting for results from I-Tasser on the mutated TAQ sequence.
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