Haynes Lab:Notebook/Engineering PC-TFs/2015/04/20: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
(Autocreate 2015/04/20 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==
*  
* Restriction Digest/Gel Verification, Concentration Reading<br>


'''Digest and Gel Verification'''
<br>
{| {{table}} border="1" cellspacing="5"
|- valign="top"
| bgcolor=#cfcfcf | Reagent
| bgcolor=#cfcfcf | Volume
| rowspan="9" |
<u><b>Expected:</b></u>
<br>
<u>BL01 Ligation Colonies [XbaI/ApaLI]</u><br>
Forward insertion: 255, 288, 883, 1243, 2037, 3017 <br>
Reverse insertion: 288, 255, 1122, 1243, 2037, 2783 <br>
No insertion: 268, 883, 1243, 2783 <br> <br>
<u>BL09 Ligation Colonies [XbaI/ApaLI]</u><br>
Forward insertion: 255, 288, 883, 1243, 3977 <br>
Reverse insertion: 255, 288, 1243, 2082, 2783 <br>
No insertion: 268, 883, 1243, 2783 <br> <br>
| rowspan="10" | [[Image:4-13-15.jpg|400px|]]<br>
15 μL/lane; 1% agarose;<br>
<u>Observed lengths:</u> <br>
W1: Ladder <br>
W2: Bl09 Lig 2 <br>
W3: Bl09 TL V1 <br>
W4: BL09 Lig 5 <br>
W5: "Bl01 LCR V3" <br>
W6: "BL01 LSR V4" <br>
W7: 1 LCR <br>
W8: Bl01 LCR V1 <br>
[http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
|-
| DNA|| 2.5 μL  2.5 μL  2.0 μL  1.0 μL  1.0 μL  3.0 μL  2.0 μL (From ladder 2 to ladder 8)
|-
| 10X buffer || 2.0 μL (for all)
|-
| XbaI || 1.0 μL (for all)
|-
| ApaLI|| 1.0 μL  (for all)
|-
| dH<sub>2</sub>O || 8.5 μL  8.5 μL  9.5 μL  9.5 μL  10.5 μL  10.5 μL
|-
| Total || 15 μL --> 37°C/ 15 min.
|}
<br>
Note: Too much buffer was used. Well 6 was weird. Most of the wells look faint.


<!-- ##### 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 19:02, 18 April 2015



 		<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>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Summary

  • Restriction Digest/Gel Verification, Concentration Reading

Digest and Gel Verification

Reagent Volume

Expected:
BL01 Ligation Colonies [XbaI/ApaLI]
Forward insertion: 255, 288, 883, 1243, 2037, 3017
Reverse insertion: 288, 255, 1122, 1243, 2037, 2783
No insertion: 268, 883, 1243, 2783

BL09 Ligation Colonies [XbaI/ApaLI]
Forward insertion: 255, 288, 883, 1243, 3977
Reverse insertion: 255, 288, 1243, 2082, 2783
No insertion: 268, 883, 1243, 2783


15 μL/lane; 1% agarose;
Observed lengths:
W1: Ladder
W2: Bl09 Lig 2
W3: Bl09 TL V1
W4: BL09 Lig 5
W5: "Bl01 LCR V3"
W6: "BL01 LSR V4"
W7: 1 LCR
W8: Bl01 LCR V1

Ladder

DNA 2.5 μL 2.5 μL 2.0 μL 1.0 μL 1.0 μL 3.0 μL 2.0 μL (From ladder 2 to ladder 8)
10X buffer 2.0 μL (for all)
XbaI 1.0 μL (for all)
ApaLI 1.0 μL (for all)
dH2O 8.5 μL 8.5 μL 9.5 μL 9.5 μL 10.5 μL 10.5 μL
Total 15 μL --> 37°C/ 15 min.


Note: Too much buffer was used. Well 6 was weird. Most of the wells look faint.


Retrieved from "https://openwetware.org/mediawiki/index.php?title=Haynes_Lab:Notebook/Engineering_PC-TFs/2015/04/20&oldid=880916"