Difference between revisions of "Team:UCLA/Notebook/Recombinant Expression"

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Revision as of 00:28, 10 July 2015

iGEM UCLA




Recombinant Silk Functionalization

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mm/dd: Entry title/short description (most recent days on top)

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mm/dd: Sample Entry

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:

  1. Tried it with newly bought ligase
  2. Left reaction overnight instead of 2 hrs
  3. 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
M T W T F S S
    1 2 3 4 5
6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30 31
August
M T W T F S S
          1 2
3 4 5 6 7 8 9
10 11 12 13 14 15 16
17 18 19 20 21 22 23
24 25 26 27 28 29 30
31
September
M T W T F S S
  1 2 3 4 5 6
7 8 9 10 11 12 13
14 15 16 17 18 19 20
21 22 23 24 25 26 27
28 29 30