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Team:RMIT Australia/Notebook
From 2010.igem.org
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<div id=" .2F15_September_2010">'''15 September''' | <div id=" .2F15_September_2010">'''15 September''' | ||
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| + | '''Mutagenesis Quikchange XL kit''' | ||
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| + | *Template Concentration = 8ng/µl | ||
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| + | *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 | ||
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| + | First mutagenesis was to turn the -35 site in the promoter region into an Afi II restriction site. | ||
| + | Dpn1 digest (2 µl) for 2hrs | ||
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<div id=" .2F15_September_2010">'''17 September''' | <div id=" .2F15_September_2010">'''17 September''' | ||
Revision as of 06:11, 23 September 2010
Notebook
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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
We optimised for low 260/280
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
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