User:Wesley J. Houston/Notebook/MAP/2013/03/18: Difference between revisions

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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;"> Project name</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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==3/18/2013==
==3/18/2013==
* Retrying ligation of KAH201(S/P)/3403 + KAH182(X/P)/1675
* Retrying ligation of KAH201(S/P)/3403 + KAH182(X/P)/1675
* Concentrations: KAH201 = 48.1 ng/uL, KAH1182 = 6.3 ng/uL
** Concentrations were determined on 2/18/13


> Ligations, following corrected version from 3/4/13, with 50 ng of vector instead of 25 ng.
> Ligations, following corrected version from 3/4/13, with 50 ng of vector instead of 25 ng.
{| class="wikitable" border="0" cellspacing="3" <!-- Ligations table -->
| <u>Ligation</u> || <font color="blue"><u>Plate results (lig : neg crtl)</u> 03/18/13</font>
|-
| 1. KAH201 (S/P)/<font color="blue">3403</font>, 50 ng + KAH182 (X/P)/1675, 50 ng || <font color="blue">TBD</font>
|-
| 2. KAH201 (S/P)/3403, 50 ng || TBD
|}
{| class="wikitable" border="0" cellspacing="3" <!-- Ligation rxn table -->
{| class="wikitable" border="0" cellspacing="3" <!-- Ligation rxn table -->
| &nbsp;            || 1    || 2    ||
| &nbsp;            || 1    || 2    ||
|-
|-
| Insert DNA (KAH182)|| 6.0  || ---  ||
| Insert DNA (KAH182)|| 8.0  || ---  ||
|-
|-
| Vector DNA (KAH201)|| 1.0  || 1.0  ||
| Vector DNA (KAH201)|| 1.0  || 1.0  ||
|-
|-
| 2x lgn buf (Roche) || 5.0  || 5.0  ||
| 2x lgn buf (Roche) || 10.0  || 5.0  ||
|-
|-
| T4 ligase (NEB)    || 1.0  || 1.0  ||
| T4 ligase (NEB)    || 1.0  || 1.0  ||
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| dH<sub>2</sub>O    ||  0  || 3.0  ||
| dH<sub>2</sub>O    ||  0  || 3.0  ||
|-
|-
| &nbsp;            || 13 μL || 10 μL ||
| &nbsp;            || 20 μL || 10 μL ||
|}
|}


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* Add 30 uL DH5α-Turbo to each ligation; plate on 100 μg/mL Amp agar
* Add 30 uL DH5α-Turbo to each ligation; plate on 100 μg/mL Amp agar


 
{| class="wikitable" border="0" cellspacing="3" <!-- Ligations table -->
| <u>Ligation</u> || <font color="blue"><u>Plate results (lig : neg crtl)</u> 03/18/13</font>
|-
| 1. KAH201 (S/P)/<font color="blue">3403</font>, 50 ng + KAH182 (X/P)/1675, 50 ng || <font color="blue">0</font>
|-
| 2. KAH201 (S/P)/3403, 50 ng || 0
|}


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Latest revision as of 22:33, 26 September 2017

Project name Main project page
Previous entry      Next entry

3/18/2013

  • Retrying ligation of KAH201(S/P)/3403 + KAH182(X/P)/1675
  • Concentrations: KAH201 = 48.1 ng/uL, KAH1182 = 6.3 ng/uL
    • Concentrations were determined on 2/18/13

> Ligations, following corrected version from 3/4/13, with 50 ng of vector instead of 25 ng.

  1 2
Insert DNA (KAH182) 8.0 ---
Vector DNA (KAH201) 1.0 1.0
2x lgn buf (Roche) 10.0 5.0
T4 ligase (NEB) 1.0 1.0
dH2O 0 3.0
  20 μL 10 μL
  • Follow quick transformation protocol
  • Add 30 uL DH5α-Turbo to each ligation; plate on 100 μg/mL Amp agar
Ligation Plate results (lig : neg crtl) 03/18/13
1. KAH201 (S/P)/3403, 50 ng + KAH182 (X/P)/1675, 50 ng 0
2. KAH201 (S/P)/3403, 50 ng 0


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