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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

Attempt 1:

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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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