Team:Calgary/2 July 2009

From 2009.igem.org

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Verification digest to verify the presence of restriction sites in the psB1AC3, psB1AK3 vectors as well as in our TOPO TA vector with LuxOD47E.
Verification digest to verify the presence of restriction sites in the psB1AC3, psB1AK3 vectors as well as in our TOPO TA vector with LuxOD47E.
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*Protocol:
*Protocol:
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5 reactions for psB1AC3 vector, 5 reactions for psB1AK3 vector, 1 reaction for gene of interest (LuxOD47E in pCR2.1-TOPO vector).
5 reactions for psB1AC3 vector, 5 reactions for psB1AK3 vector, 1 reaction for gene of interest (LuxOD47E in pCR2.1-TOPO vector).
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BBK Vector Reaction List
BBK Vector Reaction List
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Reaction 1- Not1 + React 3 Buffer
Reaction 1- Not1 + React 3 Buffer

Revision as of 19:33, 29 July 2009

University of Calgary

UNIVERSITY OF CALGARY



THIS MONTH

July
MTWTFSS
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NOTEBOOK PAGE INDEX



CALENDAR

May
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June
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29 30


July
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August
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31


September
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October
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JULY 2, 2009


CAROL

Descriptive Title of What You’re Doing

WIKI CODING HERE

CHINMOYEE

Descriptive Title of What You're Doing

WIKI CODING HERE


EMILY

Descriptive Title of What You're Doing

  • Protocol: Used GenElute Plasmid Miniprep Kit (Sigma), lot # 048k6062, used as according to the manufacturers instructions, eluting in 50 μL ddH2O.
  • Results:
Plasmid 260/280 260/230 Concentration [ng/μL]
psB1AC3 1 1.96 3.48 76.9
psB1AC3 2 1.93 2.29 98.1
psB1AC3 3 1.87 1.88 48.5
psB1AC3 4 1.90 2.11 118.9
psB1AK3 1 2.05 2.42 134.4
psB1AK3 2 1.95 2.23 55.6
psB1AK3 3 1.91 2.22 97.9
psB1AK3 4 2.03 1.92 44.5


Verification digest to verify the presence of restriction sites in the psB1AC3, psB1AK3 vectors as well as in our TOPO TA vector with LuxOD47E.

  • Protocol:


5 reactions for psB1AC3 vector, 5 reactions for psB1AK3 vector, 1 reaction for gene of interest (LuxOD47E in pCR2.1-TOPO vector).

BBK Vector Reaction List

Reaction 1- Not1 + React 3 Buffer Reaction 2- EcoRI + SpeI + React Buffer 4 Reaction 3- XbaI + PstI + React 2 Buffer Reaction 4- EcoRI + PstI + React 2 Buffer Reaction 5- XbaI + SpeI + React 4 Buffer
BBK vector tube preparation 8 uL Plasmid (psB1AC3 or psB1AK3) 2 uL appropriate React Buffer 1 uL of each appropriate restrcition enzyme ddH20 up to 20 uL
TOPO vector tube preparation 10 uL DNA (LuxOD47E in pCR2.1- TOPO vector) 7 uL ddH20 2 uL React 3 Buffer 1 uL NotI restriction enzymes
Left to digest overnight in waterbath at 37°C.




FAHD

Descriptive Title of What You're Doing

WIKI CODING HERE


IMAN

Descriptive Title of What You're Doing

WIKI CODING HERE


JAMIE

Descriptive Title of What You're Doing

WIKI CODING HERE


JEREMY

Descriptive Title of What You're Doing

WIKI CODING HERE


KATIE

Descriptive Title of What You're Doing

WIKI CODING HERE


KEVIN

Descriptive of What You're Doing

WIKI CODING HERE


MANDY

Descriptive Title of What You're Doing

WIKI CODING HERE


PATRICK

Descriptive Title of What You're Doing

WIKI CODING HERE


PRIMA

Descriptive Title of What You're Doing

WIKI CODING HERE


STEFAN

Descriptive Title of What You're Doing

WIKI CODING HERE


VICKI

Descriptive Title of What You're Doing

WIKI CODING HERE