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Team:Cambridge-JIC/Constructs/gal4 hap1gr H2RBmRFP
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Revision as of 15:44, 30 July 2014
Gal4 - Hap1GR - H2BmRFP
Design
Aim of the construct
- We'll try out 35s->GAL4->HAP1GR->H2BmRFP.
- Since H2BmRFP has not been tried in Mp, we'll also try 35s->H2BmRFP. A positive control for lab technique will be 35s->Venus N7
- We'll also use FF's original plasmid to try 35s-Hap1GR-mRFP, as a control for the Hap1GR. I expect low transformation rate as in the original plasmid, we're using 35s as promoter for HygR selection marker.
- Not sure whether 35s->Gal4 has been tried (check with BP), so 35s-Gal4-venus n7 is doable with BP's construct as a control
- As another control, we'll try 35s-Gal4-H2mRFP
End goal plasmid
Experimentation (up to date)
PCR
Attempt 1:- Start date/time: 24/07, 11:08
- End date/time:24/07, 1550
- UIDs of primers/plasmids used:
- A1, P33, P34
- A1, P35, P36
- A2, P37, P38
- A2, P39, P40
- PCR Spreadsheet (on google drive)
- Gel 27/07/2014
Gibson
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E-Coli transformation
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Agrobacteria transformation
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Spore preparation
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Spore transformation
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Evaluation
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