Team:RMIT Australia/Notebook

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Notebook

May
MTWTFSS
          [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F1_May_2010 1] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F2_May_2010 2]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F3_May_2010 3] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F4_May_2010 4] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F5_May_2010 5] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F6_May_2010 6] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F7_May_2010 7] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F8_May_2010 8] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F9_May_2010 9]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F10_May_2010 10] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F11_May_2010 11] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F12_May_2010 12] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F13_May_2010 13] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F14_May_2010 14] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F15_May_2010 15] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F16_May_2010 16]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F17_May_2010 17] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F18_May_2010 18] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F19_May_2010 19] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F20_May_2010 20] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F21_May_2010 21] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F22_May_2010 22] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F23_May_2010 23]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F24_May_2010 24] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F25_May_2010 25] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F26_May_2010 26] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F27_May_2010 27] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F28_May_2010 28] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F29_May_2010 29] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F30_May_2010 30]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F31_May_2010 31]
June
MTWTFSS
  [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F1_June_2010 1] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F2_June_2010 2] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F3_June_2010 3] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F4_June_2010 4] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F5_June_2010 5] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F6_June_2010 6]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F7_June_2010 7] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F8_June_2010 8] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F9_June_2010 9] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F10_June_2010 10] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F11_June_2010 11] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F12_June_2010 12] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F13_June_2010 13]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F14_June_2010 14] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F15_June_2010 15] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F16_June_2010 16] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F17_June_2010 17] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F18_June_2010 18] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F19_June_2010 19] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F20_June_2010 20]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F21_June_2010 21] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F22_June_2010 22] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F23_June_2010 23] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F24_June_2010 24] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F25_June_2010 25] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F26_June_2010 26] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F27_June_2010 27]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F28_June_2010 28] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F29_June_2010 29] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F30_June_2010 30]
July
MTWTFSS
      [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F1_July_2010 1] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F2_July_2010 2] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F3_July_2010 3] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F4_July_2010 4]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F5_July_2010 5] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F6_July_2010 6] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F7_July_2010 7] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F8_July_2010 8] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F9_July_2010 9] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F10_July_2010 10] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F11_July_2010 11]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F12_July_2010 12] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F13_July_2010 13] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F14_July_2010 14] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F15_July_2010 15] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F16_July_2010 16] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F17_July_2010 17] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F18_July_2010 18]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F19_July_2010 19] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F20_July_2010 20] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F21_July_2010 21] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F22_July_2010 22] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F23_July_2010 23] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F24_July_2010 24] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F25_July_2010 25]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F26_July_2010 26] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F27_July_2010 27] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F28_July_2010 28] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F29_July_2010 29] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F30_July_2010 30] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F31_July_2010 31]
August
MTWTFSS
            [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F1_August_2010 1]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F2_August_2010 2] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F3_August_2010 3] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F4_August_2010 4] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F5_August_2010 5] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F6_August_2010 6] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F7_August_2010 7] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F8_August_2010 8]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F9_August_2010 9] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F10_August_2010 10] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F11_August_2010 11] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F12_August_2010 12] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F13_August_2010 13] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F14_August_2010 14] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F15_August_2010 15]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F16_August_2010 16] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F17_August_2010 17] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F18_August_2010 18] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F19_August_2010 19] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F20_August_2010 20] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F21_August_2010 21] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F22_August_2010 22]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F23_August_2010 23] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F24_August_2010 24] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F25_August_2010 25] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F26_August_2010 26] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F27_August_2010 27] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F28_August_2010 28] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F29_August_2010 29]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F30_August_2010 30] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F31_August_2010 31]
September
MTWTFSS
    [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F1_September_2010 1] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F2_September_2010 2] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F3_September_2010 3] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F4_September_2010 4] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F5_September_2010 5]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F6_September_2010 6] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F7_September_2010 7] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F8_September_2010 8] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F9_September_2010 9] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F10_September_2010 10] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F11_September_2010 11] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F12_September_2010 12]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F13_September_2010 13] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F14_September_2010 14] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F15_September_2010 15] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F16_September_2010 16] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F17_September_2010 17] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F18_September_2010 18] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F19_September_2010 19]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F20_September_2010 20] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F21_September_2010 21] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F22_September_2010 22] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F23_September_2010 23] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F24_September_2010 24] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F25_September_2010 25] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F26_September_2010 26]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F27_September_2010 27] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F28_September_2010 28] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F29_September_2010 29] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F30_September_2010 30]
October
MTWTFSS
        [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F1_October_2010 1] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F2_October_2010 2] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F3_October_2010 3]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F4_October_2010 4] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F5_October_2010 5] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F6_October_2010 6] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F7_October_2010 7] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F8_October_2010 8] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F9_October_2010 9] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F10_October_2010 10]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F11_October_2010 11] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F12_October_2010 12] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F13_October_2010 13] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F14_October_2010 14] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F15_October_2010 15] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F16_October_2010 16] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F17_October_2010 17]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F18_October_2010 18] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F19_October_2010 19] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F20_October_2010 20] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F21_October_2010 21] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F22_October_2010 22] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F23_October_2010 23] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F24_October_2010 24]
[http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F25_October_2010 25] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F26_October_2010 26] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F27_October_2010 27] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F28_October_2010 28] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F29_October_2010 29] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F30_October_2010 30] [http://2010.igem.org/Team:RMIT_Australia/Notebook#.2F31_October_2010 31]


28 June

Did OH&S training and PC2 training

29 June

Began research on what we were going to use to promote taq

30 June

Looked at T7 with a Lac operon as a promotore of taq

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

14 July

Looked at prices of Hotels in Boston

19 July

began looking at the structure of taq, its thermostability began looking at different mutations that can be done to taq to increase its thermostability without changing its structure

21 July

sent in sequence of wild type taq to iTasser

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

19 August

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. Went into the lab and made SOC media and LB- agar plates

20 August

Did two PCR reaction and a transformation The first PCR we ran was a touch down PCR. Temperatures ranged from 69 to 64. The second PCR was a two step PCR.

30 August

Ordered the TAQ polymerase from Mr. Gene. Should have it in 15 days. Meeting today at 10am to discuss project budget and costings. After the meeting we all are sitting together for a writing session.

31 August

Spent the morning organising our flights In the afternoon we all sat together for a writing session

1 September

We all sat together for another writing session.

7 September

Had a team meeting

14 September

Had a team meeting and made a lab time table, we also set out tasks which need to be done by the next meeting - Update the wiki

- Finalise the form for the ethics committee for pennstate

- Call up possible sponsors

- Collect Tshirts

- Work on abstract and team Roster

- research heat liable bonds

- order peptides

- chat with emilio about possible bio assays

- work on making the promotor part

15 September

Mutagenesis Quikchange XL kit

  • Template Concentration = 8ng/µl
  • Fwd 77ng/µl (low 260/280)
  • Rev 86.5 ng/µl (low 260/280)

We optimised for low 260/280

  • 5 µl buffer
  • 1.9 µl template
  • 1.7 µl Fwd
  • 1.62 µl Rev
  • 1 µl dNTP
  • 38.78 µl H2O --> reaction volume 50 µl
  • 1 µl Pfu

First mutagenesis was to turn the -35 site in the promoter region into an Afi II restriction site. Dpn1 digest (2 µl) for 2hrs


17 September

- finalized project abstract

19 September


- sent off project abstract to HQ
Retrieved from "http://2010.igem.org/Team:RMIT_Australia/Notebook"