iGEM UCLA

Recombinant Silk Functionalization

=Raw lab notebook entries= {{#calendar: year=2015 | month= jul | title = Team:UCLA/Notebook/Recombinant_Expression | query=preload=Template:UCLA}} {{#calendar: year=2015 | month= aug | title = Team:UCLA/Notebook/Recombinant_Expression | query=preload=Template:UCLA}} {{#calendar: year=2015 | month= sep | title = Team:UCLA/Notebook/Recombinant_Expression | query=preload=Template:UCLA}}

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Today we began cloning our GFP.

PCR'd off template
Ran gel
Restriction digest
Ligated into backbone

For our ligation, we made the following modifications:

Tried it with newly bought ligase
Left reaction overnight instead of 2 hrs
Vector to insert ratio was 1:5 instead of 1:3

PCR Reaction:

Component	Volume
5X Q5 Reaction Buffer	5
10 mM dNTPs	0.5
10 uM Forward (primer 3/7)	1.25
10 uM Reverse (primer 8)	1.25
Template (diluted to 1ng/uL)	0.5
Q5 High Fidelity DNA Polymerase	0.25
Nuclease Free Water	16.25

Gel: Lot of bands, all at correct sizes

Goals

Julian's Goals

Finish Purification of ABD containing construct
Create hydrogel with ABD and B.mori silk and see if it retains/binds albumin

Achievements

Julian's Achievements

Cloned the ABD construct and begun purification methods

What to accomplish next

Julian

Test albumin binding properties of the ABD silk construct
test hydrogel formation and albumin retention

Raw lab notebook entries

July
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August
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September
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Team:UCLA/Notebook/Recombinant Expression