Haynes Lab:Notebook/CRISPR Editing/2014/08/07: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
(fix raw html notebook nav) |
|||
(7 intermediate revisions by one other user not shown) | |||
Line 2: | Line 2: | ||
|- | |- | ||
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
|- | |- | ||
| colspan="2"| | | colspan="2"| | ||
Line 54: | Line 54: | ||
| mCherry gRNA4 || CRISPR 4d || " || 329 || ~180, 149 | | mCherry gRNA4 || CRISPR 4d || " || 329 || ~180, 149 | ||
|- | |- | ||
| mCherry || | | mCherry || mock || " || 329 || no cutting | ||
|- | |- | ||
| luciferase gRNA20 || g20 +dox || gRNA 021 US P F / gRNA 027 ES P R || 1295 || ~165, 1130 | | luciferase gRNA20 || g20 +dox || gRNA 021 US P F / gRNA 027 ES P R || 1295 || ~165, 1130 | ||
Line 108: | Line 108: | ||
| -dox || 1.169 || 1.8 || 58.5 | | -dox || 1.169 || 1.8 || 58.5 | ||
|- | |- | ||
| | | U2OS mock || --- || --- || not done yet | ||
|} | |} | ||
'''Mismatch annealing''' (continued on 08/07/14) | |||
* Zhang Lab approach - do not mix with a wild type reference, since ~50% or more of the amplicons in the mutated cell sample will be wild type anyway | |||
** 200 ng clean PCR, 1x HF Phusion buffer, in final volume of 20 μL | |||
# CRISPR 1d | |||
# CRISPR 2d | |||
# CRISPR 4d | |||
# g20 +dox | |||
# g20 -dox | |||
# g29 +dox | |||
# | # g29 -dox | ||
# | # +dox | ||
# -dox | |||
# U2OS mock | |||
Line 142: | Line 146: | ||
** 4 ºC/ hold ∞ | ** 4 ºC/ hold ∞ | ||
'''SURVEYOR nuclease reaction''' | |||
Latest revision as of 00:10, 27 September 2017
Project name | Main project page Previous entry Next entry | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
08/07/2014
CRISPR on chromatin: transfections
Luc-mCh swap: building the donor sequence
SURVEYOR assay
|