Team:Cornell/Week 9
From 2011.igem.org
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(→Thursday, August 4) |
(→Thursday, August 4) |
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==Thursday, August 4== | ==Thursday, August 4== | ||
Afternoon lab work done by: Claire Paduano, Charlie Chung | Afternoon lab work done by: Claire Paduano, Charlie Chung | ||
+ | |||
+ | :'''Objective''' | ||
+ | ::Ligation of GFP/RFP/VioE + avitag + backbone | ||
+ | :'''Ligation''' | ||
+ | ::*''GFP + Avi-Tag + Backbone'' | ||
+ | ::::10.6μL pZE12 +avitag vector backbone | ||
+ | ::::3.6μL GFP | ||
+ | ::::2.9μL H2O | ||
+ | ::::2.0μL T4 DNA Ligase Buffer | ||
+ | ::::<u>1.0μL T4 DNA Ligase</u> | ||
+ | ::::20μL Total | ||
+ | ::*''RFP + Avi-Tag + Backbone'' | ||
+ | ::::10.3μL pZE12 + avitag vector backbone | ||
+ | ::::6.7μL RFP | ||
+ | ::::2.0μL T4 DNA Ligase Buffer | ||
+ | ::::<u>1.0μL T4 DNA Ligase</u> | ||
+ | ::::20μL Total | ||
+ | ::*''VioE + Avi-Tag + Backbone'' | ||
+ | ::::2.3μL H2O | ||
+ | ::::4.1μL VioE | ||
+ | ::::10.6μL pZE12 + avitag vector backbone | ||
+ | ::::2.0μL T4 DNA Ligase Buffer | ||
+ | ::::<u>1.0μL T4 DNA Ligase</u> | ||
+ | ::::20μL Total | ||
+ | ::In all above reactions, volume of plasmid vector added corresponds with 100ng backbone. | ||
+ | ::Prepare control ligation reactions (inserts replaced with H2O) for each of the above constructs. | ||
+ | :::*Same volume of vector backbone, buffer, ligase | ||
+ | :::*Volumes of all inserts (i.e. gene or primer pairs) go toward volume of H2O | ||
+ | ::After ligation setup, incubate in 16°C waterbath overnight. | ||
==Friday== | ==Friday== |
Revision as of 20:50, 4 August 2011
Week 1 | Week 2 - 5 | Week 6 | Week 7 | Week 8 | Week 9 | Week 10 | Week 11 | Week 12 | Week 13 | Week 14 | Week 15 | Week 16 | Week 17 | Week 18 | Week 19 | Week 20 | Week 21 |
July 31st - August 6th
Sunday
Monday
Tuesday
Wednesday
Thursday, August 4
Afternoon lab work done by: Claire Paduano, Charlie Chung
- Objective
- Ligation of GFP/RFP/VioE + avitag + backbone
- Ligation
- GFP + Avi-Tag + Backbone
- 10.6μL pZE12 +avitag vector backbone
- 3.6μL GFP
- 2.9μL H2O
- 2.0μL T4 DNA Ligase Buffer
- 1.0μL T4 DNA Ligase
- 20μL Total
- RFP + Avi-Tag + Backbone
- 10.3μL pZE12 + avitag vector backbone
- 6.7μL RFP
- 2.0μL T4 DNA Ligase Buffer
- 1.0μL T4 DNA Ligase
- 20μL Total
- VioE + Avi-Tag + Backbone
- 2.3μL H2O
- 4.1μL VioE
- 10.6μL pZE12 + avitag vector backbone
- 2.0μL T4 DNA Ligase Buffer
- 1.0μL T4 DNA Ligase
- 20μL Total
- In all above reactions, volume of plasmid vector added corresponds with 100ng backbone.
- Prepare control ligation reactions (inserts replaced with H2O) for each of the above constructs.
- Same volume of vector backbone, buffer, ligase
- Volumes of all inserts (i.e. gene or primer pairs) go toward volume of H2O
- After ligation setup, incubate in 16°C waterbath overnight.