Haynes Lab:Notebook/Engineering PC-TFs/2014/08/08: Difference between revisions
(Autocreate 2014/08/08 Entry for Haynes_Lab:Notebook/Engineering_PC-TFs) |
|||
| Line 7: | Line 7: | ||
<!-- ##### 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== | ||
* | *Concentration reading for gel purified PcTFs, ligation product minipreps. | ||
*Gel verification of ligation attempt 3. | |||
---- | |||
'''Concentration Readings''' | |||
<br> | |||
{| {{table}} border="1" cellspacing="3" <!-- Plates. table --> | |||
|- valign="top" | |||
| rowspan="7" | [[Image:Bl01,5,9 gel p conc.jpg|400px|Hover name]] | |||
|} | |||
<br> | |||
---- | |||
'''Gel Verification''' | |||
{| {{table}} border="1" cellspacing="5" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="9" | | |||
<u><b>Expected:</b></u> | |||
<br> | |||
<br>BL01(Well: 2), BL05(Well:3-6) <br> | |||
Forward insertion: 710, 7010. <br> | |||
Reverse insertion: 3230, 4490. <br> | |||
No insertion: 710, 4490. <br><br> | |||
<u>CMV/MV9 [XbaI/NruI] (Well: 7)</u> <br> | |||
710, 4490<br><br> | |||
<u>BL09 Ligation Colonies 1-6[XbaI/NruI] (Well: 8-13)</u><br> | |||
Forward insertion: 710, 5921 <br> | |||
Reverse insertion: 2141, 4490 <br> | |||
No insertion: 710, 4490<br> <br> | |||
| rowspan="10" | [[Image:08 08 14 pctf lig3 ver20140808 141240.jpg|400px|Today's gel]]<br> | |||
15 μL/lane; 1% agarose;<br> | |||
<u>Observed lengths:</u> <br> | |||
W2(BL01Lig1): 5000, 700; <br> | |||
W3(BL05Lig1): 5000, 700; <br> | |||
W4(BL05Lig2): 5000, 700; <br> | |||
W5(BL05Lig3): 5000, 700; <br> | |||
W6(BL05Lig4): 5000, 700; <br> | |||
W7(Empty CMV/MV9): 5000, 700; <br> | |||
W8(BL09Lig1): 5000, 700; <br> | |||
W9(BL09Lig2): 7000, 700; <br> | |||
W10(BL09Lig3): 5000, 700; <br> | |||
W11(BL09Lig4): 5000, 700; <br> | |||
W12(BL09Lig5): 7000, 700;<br> | |||
W13(BL09Lig6): 5000, 700; <br> | |||
[http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | |||
|- | |||
| DNA(BL01x2, BL05x1, BL09x2, CMV/MV9x1) || 2.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 1.0 μL | |||
|- | |||
| NruI || 1.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 9.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br> | |||
BL09Lig2&5 sent in for sequencing. | |||
<!-- ##### 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. ##### --> | ||
Revision as of 14:26, 8 August 2014
  |
<html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | |||||||||||||||||
Summary
Concentration Readings
Gel Verification
| ||||||||||||||||||
