IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/06/28: Difference between revisions

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! NEB Buffer 3 (10x)
! NEB Buffer 3 (10x)
| 2 || 2 || 2 || 2
| 2 || 2 || 2 || 2
| -
|-
! diH<sub>2</sub>O
! diH<sub>2</sub>O
| 10 || 7 || 2 || 7
| 10 || 7 || 2 || 7
|
|}
|}


Revision as of 11:57, 28 June 2010

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Entry title

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PCR Reactions

Mix of Parts 1,2,3 w/ primers A&B (simultaneous PCR) 3 reactions (GFP; Bet; LTP)

Annealing Temp: 60C Extension Time: 15 sec

Name Amount (uL)
10mM dNTPs 1
5x Phusion Buffer 10
Phusion Polymerase .5
DNA (Parts "1" "2" "3") 1(parts 1&2); .5 (part 3)= 2.5 total
AB primer mix (1x) 2.5
Water 35


2 reactions (GFP, LTP)

Name Amount (uL)
10mM dNTPs 1
5x Phusion Buffer 10
Phusion Polymerase .5
DNA (Parts "1" "2" "3") 1(each)= 3 total
AB primer mix (1x) 2.5
Water 35


Mix of Parts 1&2 w/ Primers A&2 3 reactions (GFP; Bet; LTP)

Mix of Parts 1&2 & 3 w/ Primers A&B

Gels

Team Flavour

Miniprep of Miraculin and Brazzein Constructs

  • Protocol used was from the Qiagen Miniprep Kit
Nanodrop Specs
Name Concentration (ng/μL)
Miraculin 1 159.6
Miraculin 2 182.2
Brazzein 1 91.2
Brazzein 2 33.7

DTL (Digestion, Ligation, Transformation) of The Big Three (pENTCUP2, NOSt, NOSt + STOP)

Digestion Reaction
pENTCUP2 NOSt NOSt+STOP B15
DNA 4 7 12 7
NEB Buffer 3 (10x) 2 2 2 2
diH2O 10 7 2 7



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