User:Daniel A. Vargas/Notebook/General lab notebook/2015/08/03: Difference between revisions

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{|{{table}} width="800"
{|{{table}} width="800"
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|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;"> Open Chromatin</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|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]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
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==Quality Check of PCR and RED==
==Quality Check of PCR and RED==
MV10 digest
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Reagent'''
| align="center" style="background:#f0f0f0;"|'''Volume'''
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| MV10 |||15.00
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| Xba1 |||1.00
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| MBN|||1.00
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| 10X FD buffer |||3.00
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| dH<sub>2</sub>O||| 10.00
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| Total  |||30.00
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Run at 37<sup>o</sup>C for 15 min
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| bgcolor=#cfcfcf | Volume
| bgcolor=#cfcfcf | Volume
| rowspan="7" | <u>Expected:</u><br>1. MV10 = 5191, <br>2. ATF2 = 906 <br>3. ATF2 neg ctrl = 0<br>4. Gal4db/mCherry = 1170<br>5. Gal4db/mCherry = 0<br>
| rowspan="7" | <u>Expected:</u><br>1. MV10 = 5191, <br>2. ATF2 = 906 <br>3. ATF2 neg ctrl = 0<br>4. Gal4db/mCherry = 1170<br>5. Gal4db/mCherry = 0<br>
| rowspan="7" | [[image: Plasmidverification.jpg|200px|Damaged Gel]]<br>10 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
| rowspan="7" | [[image: QC_of_pcr_red.jpg|200px|Damaged Gel]]<br>10 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
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| DNA || 3.0 μL
| DNA || 3.0 μL

Latest revision as of 01:04, 27 September 2017

Open Chromatin Main project page
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Quality Check of PCR and RED

MV10 digest

Run at 37oC for 15 min
Reagent Volume
MV10 15.00
Xba1 1.00
MBN 1.00
10X FD buffer 3.00
dH2O 10.00
Total 30.00
Reagent Volume Expected:
1. MV10 = 5191,
2. ATF2 = 906
3. ATF2 neg ctrl = 0
4. Gal4db/mCherry = 1170
5. Gal4db/mCherry = 0
 Damaged Gel
10 μL/lane; 1% agarose; Ladder
DNA 3.0 μL
6X BioLabs purple dye 3.0 μL
dH2O 4.0 μL
  10 μL

Qiagen clean-up Results

Plasmid Conc. (ng/µL) St Dev (ng/µL) 260/280
MV10 46.37 1.13 1.75
ATF2 64.350 2.15 1.90
Gal4DB/mCherry 50.873 1.13 1.93


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