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

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===Miniprep of Miraculin and Brazzein Constructs===
===Miniprep of Miraculin and Brazzein Constructs===
Nanodrop Specs:
{| border ="1"
{| class="wikitable" border ="1px solid black"
|+Nanodrop Specs
|-
|-
!Name  
!Name  
Line 78: Line 78:
|-
|-
|Miraculin 1
|Miraculin 1
|159.6
| align="right"| 159.6
|-
|-
|Miraculin 2
|Miraculin 2
|182.2
| align="right" | 182.2
|-
|-
|Brazzein 1
|Brazzein 1
|91.2
| align="right" | 91.2
|-
|-
|Brazzein 2
|Brazzein 2
|33.7
| align="right" | 33.7
|-
|-
|}
|}
Miraculin 1: 159.6 ng/μL


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

Revision as of 11:46, 28 June 2010

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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

Team Flavour

Miniprep of Miraculin and Brazzein Constructs

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)

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



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