Haynes Lab:Notebook/Engineering PC-TFs/2014/07/09: Difference between revisions
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<!-- ##### 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== | ||
'''Liquid Culture Minipreps''' | |||
<br> | |||
Eluted with 70μL dH<sub>2</sub>O.<br> | |||
---- | |||
{| {{table}} | |||
|- bgcolor=#cfcfcf | |||
| Plasmid || OD260 || OD260/280 || ng/μL | |||
|- | |||
| 1. BL01 C1 || 0.289 || 1.89 || 288.704 | |||
|- | |||
| 1. BL01 C2 || 0.463 || 1.918 || 462.857 | |||
|- | |||
| 1. BL01 C3 || 0.495 || 1.915 || 495.398 | |||
|} | |||
<br> | |||
{| {{table}} | |||
|- bgcolor=#cfcfcf | |||
| Plasmid || OD260 || OD260/280 || ng/μL | |||
|- | |||
| 1. BL05 C1 || 0.047 || 2.124 || 46.727 | |||
|- | |||
| 1. BL05 C2 || 0.301 || 1.912 || 301.14 | |||
|- | |||
| 1. BL05 C3 || 0.075 || 1.97 || 74.969 | |||
|} | |||
<br> | |||
{| {{table}} | |||
|- bgcolor=#cfcfcf | |||
| Plasmid || OD260 || OD260/280 || ng/μL | |||
|- | |||
| 1. BL09 C1 || 0.059 || 1.986 || 59.185 | |||
|- | |||
| 1. BL09 C2 || 0.236 || 1.878 || 235.782 | |||
|- | |||
| 1. BL09 C3 || 0.185 || 1.958 || 184.993 | |||
|} | |||
<br> | |||
---- | |||
'''Restriction Digest''' | |||
{| {{table}} border="1" cellspacing="5" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="9" | <u>Expected:</u><br>BL01(W1:2-4), BL05(W1:5, W2:2-3) <br>Forward insertion: 710, 7010. <br>Reverse insertion: 3230, 4490. <br>No insertion: 710, 4490.<br> BL09 (W2:4-6) <br>Forward insertion: 710, 5921 <br>Reverse insertion: 2141, 4490 <br>No insertion: 710, 4490.<br> *Final well is empty CMV/MV9. | |||
| rowspan="10" | [[Image:W120140709 174255.jpg|400px|Today's gel]]<br>15 μL/lane; 1% agarose;<br> <u>Observed lengths:</u> W2: 7100, 700; W3: 7100, 5200, 2500; W4: *5200, 2500, 710; W5: 5000, 2500; W6: 5000, 2500, 710.<br> *Uncut plasmid.<br> [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | |||
| rowspan="10" | [[Image:W220140709 174229.jpg|400px|Today's gel]]<br>15 μL/lane; 1% agarose;<br> <u>Observed lengths:</u> W2: *7100, 700; W3: *6100, 2500, 710; W4: *7200, 2500, 710; W5: 5000, 4000; W6: *5500, 2500, 750.<br> *Uncut plasmid.<br> [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | |||
|- | |||
| DNA(BL01x3, BL05x3, BL09x3, 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> | |||
'''Results''' | |||
<br> | |||
Top W2: BL01 possible success.<br> | |||
Bottom W2: BL05 possible success.<br> | |||
Bottom W4: BL09 possible success.<br> | |||
---- | |||
'''Sequencing Results Order 9218''' | |||
<br> | |||
<u>BL01rA<br></u> | |||
*Used 1μL BL01 miniprep 1, 1μL DD123 B primer, 8μLdH<sub>2</sub>O. | |||
*'''Results''': Matches were found after the primer and then just before CMV suggesting no insert. | |||
<br> | |||
<u>BL01rB</u><br> | |||
*Used 1μL BL01 miniprep 2, 1μL DD123 B primer, 8μLdH<sub>2</sub>O. | |||
*'''Results''': Matches were found after the primer and then just before CMV suggesting no insert. | |||
<br> | |||
<u>BL09rA</u><br> | |||
*Used 1μL BL09 miniprep 1, 1μL DD123 B primer, 8μLdH<sub>2</sub>O. | |||
*'''Results''': Matches were found after the primer and then just before CMV suggesting no insert. | |||
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__NOTOC__ | __NOTOC__ |
Latest revision as of 00:05, 27 September 2017
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SummaryLiquid Culture Minipreps
Restriction Digest
Sequencing Results Order 9218
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