Haynes Lab:Notebook/CRISPR Editing/2015/02/02: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
Line 17: Line 17:
|}
|}


''PCR of gRNA from Luc14 and GAL4-EED+dox samples, g034''
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Cell type'''
| align="center" style="background:#f0f0f0;"|'''treatment'''
| align="center" style="background:#f0f0f0;"|'''rep'''
| align="center" style="background:#f0f0f0;"|'''Concentration'''
| align="center" style="background:#f0f0f0;"|'''ul for 200ng'''
| align="center" style="background:#f0f0f0;"|'''ul for 800ng'''
|-
| Luc14||g034||1||71.774||2.79||11.15
|-
| Luc14||g034||2||117.513||1.70||6.81
|-
| Luc14||g034||3||108.009||1.85||7.41
|-
| GAL4-EED +dox||g034||1||86.013||2.33||9.30
|-
| GAL4-EED +dox||g034||2||105.127||1.90||7.61
|-
| GAL4-EED +dox||g034||3||122.48||1.63||6.53
|}


Mastermix 1
{| {{table}}
| align="center" style="background:#f0f0f0;"|''''''
| align="center" style="background:#f0f0f0;"|'''1rxn'''
| align="center" style="background:#f0f0f0;"|'''25.5'''
|-
| H20||30.8||785.4
|-
| HF 5X MasterMix||10||255
|-
| dNTPs||0.2||5.1
|-
| FP||2.5||63.75
|-
| RP||2.5||63.75
|-
| Total||46||1173
|}
Mastermix 2
{| {{table}}
| align="center" style="background:#f0f0f0;"|''''''
| align="center" style="background:#f0f0f0;"|'''1rxn'''
| align="center" style="background:#f0f0f0;"|'''4.2rxns'''
|-
| MM1||46||193.2
|-
| Template||2||8.4
|-
| DMSO||1.5||6.3
|-
| Phusion||0.5||2.1
|-
| Total||50||205
|}
Transfer 50μL of MM2 into each of 4-PCR tubes.<br>
PCR setup
PCR
:98°C for 3 min
:30 cycles
::98°C for 10s
::69.5°C for 30s
::72°C for 60s
:72°C for 10 min
:4°C forever
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
|}
|}


__NOTOC__
__NOTOC__

Revision as of 11:08, 2 February 2015

Project name <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

02/02/2015

Sample Read# 260 280 260/280 ng/µL
ryan's cloning insert 0.109 0.059 1.853 108.796

PCR of gRNA from Luc14 and GAL4-EED+dox samples, g034

Cell type treatment rep Concentration ul for 200ng ul for 800ng
Luc14 g034 1 71.774 2.79 11.15
Luc14 g034 2 117.513 1.70 6.81
Luc14 g034 3 108.009 1.85 7.41
GAL4-EED +dox g034 1 86.013 2.33 9.30
GAL4-EED +dox g034 2 105.127 1.90 7.61
GAL4-EED +dox g034 3 122.48 1.63 6.53


Mastermix 1

' 1rxn 25.5
H20 30.8 785.4
HF 5X MasterMix 10 255
dNTPs 0.2 5.1
FP 2.5 63.75
RP 2.5 63.75
Total 46 1173


Mastermix 2

' 1rxn 4.2rxns
MM1 46 193.2
Template 2 8.4
DMSO 1.5 6.3
Phusion 0.5 2.1
Total 50 205

Transfer 50μL of MM2 into each of 4-PCR tubes.

PCR setup

PCR

98°C for 3 min
30 cycles
98°C for 10s
69.5°C for 30s
72°C for 60s
72°C for 10 min
4°C forever


Retrieved from "https://openwetware.org/mediawiki/index.php?title=Haynes_Lab:Notebook/CRISPR_Editing/2015/02/02&oldid=858094"