Haynes Lab:Notebook/Engineering PC-TFs/2014/06/09: Difference between revisions
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|style="background-color: #FFCC00"|<br>[[Image:Hayneslab3.gif|200px|center]][[Image:Asu logo 3.gif|200px|center]]<span style="font-size:22px;"><br></span> | |style="background-color: #FFCC00"|<br>[[Image:Hayneslab3.gif|200px|center]][[Image:Asu logo 3.gif|200px|center]]<span style="font-size:22px;"><br></span> | ||
|style="background-color: #800000" align="center"| | |style="background-color: #800000" 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}}}}]]}} | ||
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| colspan="2"| | | colspan="2"| | ||
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | ||
==Summary== | ==Summary== | ||
'''Digest All the Things'''<br> | |||
---- | |||
'''Well One: Ladder'''<br> | |||
'''Well Two'''<br> | |||
{| {{table}} border="1" cellspacing="3" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. Unknown <br><u>Observed:</u><br> 3k, 7k, 2k<br> | |||
|- | |||
| DNA(CMV/MV9) || 5.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 0.0 μL | |||
|- | |||
| NruI || 0.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 8.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br> | |||
'''Well Three'''<br> | |||
{| {{table}} border="1" cellspacing="3" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. 5200bp <br><u>Observed:</u><br> 5.2k, 2k<br> | |||
|- | |||
| DNA(CMV/MV9) || 5.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 1.0 μL | |||
|- | |||
| NruI || 0.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 7.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|}<br> | |||
'''Well Four'''<br> | |||
{| {{table}} border="1" cellspacing="3" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. 5200bp <br><u>Observed:</u><br> 5.2k, 2k<br> | |||
|- | |||
| DNA(CMV/MV9) || 5.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 0.0 μL | |||
|- | |||
| NruI || 1.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 7.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br> | |||
'''Well Five'''<br> | |||
{| {{table}} border="1" cellspacing="3" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. 710bp, 4490bp <br><u>Observed:</u><br> 4k, 2k, 700<br> | |||
|- | |||
| DNA(CMV/MV9) || 5.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 1.0 μL | |||
|- | |||
| NruI || 1.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 6.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br>'''Well Six'''<br> | |||
{| {{table}} border="1" cellspacing="3" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. BL01 = Insert (2520bp), V0120(3265bp) <br><u>Observed:</u><br> 3.1k, 2.6k<br> | |||
|- | |||
| DNA(BL01A) || 5.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 1.0 μL | |||
|- | |||
| SpeI || 1.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 7.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br>'''Well Seven'''<br> | |||
{| {{table}} border="1" cellspacing="3" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. BL01 = 2520bp, 3265bp <br> <u>Observed:</u><br> 3.1k, 2.6k, 7k<br> | |||
|- | |||
| DNA(BL01B) || 5.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 1.0 μL | |||
|- | |||
| SpeI || 1.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 7.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br>'''Well Eight'''<br> | |||
{| {{table}} border="1" cellspacing="3" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br> 2520bp, 3265bp <u>Observed:</u><br> Faint: 3.1k, 2.6k, 7k<br> | |||
|- | |||
| DNA(BL05) || 5.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 1.0 μL | |||
|- | |||
| SpeI || 1.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 7.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br>'''Well Nine'''<br> | |||
{| {{table}} border="1" cellspacing="3" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br> 1431bp, 3265bp <u>Observed:</u><br> 3.1k, 4.9k, 1.5k <br> | |||
|- | |||
| DNA(BL09) || 5.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 1.0 μL | |||
|- | |||
| SpeI || 1.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 7.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br>'''Well 10'''<br> | |||
{| {{table}} border="1" cellspacing="3" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. BL01 = 7720bp <br> <u>Observed:</u><br> 4k, 2k, 1.5k <br> | |||
|- | |||
| DNA(CMV/MV9 & BL01) || 5.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 0.0 μL | |||
|- | |||
| NruI || 1.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 7.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br>'''Well 11'''<br> | |||
{| {{table}} border="1" cellspacing="3" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. dependent on ligation direction <br><u>Observed:</u><br>4k, 2k, 1.5k<br> | |||
|- | |||
| DNA(CMV/MV9 & BL01) || 5.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 1.0 μL | |||
|- | |||
| NruI || 1.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 6.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br> | |||
---- | |||
'''Gel Verification''' | |||
{| {{table}} border="1" cellspacing="3" <!-- Plates. table --> | |||
|- valign="top" | |||
| rowspan="7" | [[Image:20140609 172345.jpg|400px|Hover name]] <br> [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | |||
|} | |||
<!-- ##### 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. ##### --> |
Latest revision as of 00:02, 27 September 2017
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SummaryDigest All the Things Well One: Ladder
Well Four
Gel Verification
|