Haynes Lab:Notebook/Engineering PC-TFs/2015/01/12: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
Line 26: Line 26:
15 μL/lane; 1% agarose;<br>  
15 μL/lane; 1% agarose;<br>  
<u>Observed lengths:</u> <br>
<u>Observed lengths:</u> <br>
W2 BL05 LCR SV1 (small volume lcr reaction product: 4uL)
W2 BL05 LCR SV1 (small volume lcr reaction product: 4uL) <br>
W3 BL05 LCR SV2
W3 BL05 LCR SV2<br>
W4 BL05 LCR LV1 (large volume lcr reaction product: 16uL)
W4 BL05 LCR LV1 (large volume lcr reaction product: 16uL)<br>
W5 BL05 LCR LV2
W5 BL05 LCR LV2<br>
W6 BL01 LCR SV1
W6 BL01 LCR SV1<br>
W7 BL01 LCR SV2
W7 BL01 LCR SV2<br>
W8 Empty CMV/MV9
W8 Empty CMV/MV9<br>
W9 BL01 LCR LV1
W9 BL01 LCR LV1<br>
W10 BL01 LCR LV2
W10 BL01 LCR LV2<br>
W11 LCR Negative Control Colony
W11 LCR Negative Control Colony<br>
W12 BL05 Traditional Ligation SV1
W12 BL05 Traditional Ligation SV1<br>
W13 BL05 Traditional Ligation SV2
W13 BL05 Traditional Ligation SV2<br>
W14 BL05 Traditional Ligation LV1
W14 BL05 Traditional Ligation LV1<br>
[http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
[http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
|-
|-

Revision as of 16:23, 12 January 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

Ligation Digest and Gel Verification

Reagent Volume

Expected:
BL09 Ligation Colonies [XbaI/NruI]
Forward insertion: 710, 7000
Reverse insertion: 3238, 4528
No insertion: 710, 4490

CMV/MV9 with [XbaI/NruI]
710, 4490

Today's gel

15 μL/lane; 1% agarose;
Observed lengths:
W2 BL05 LCR SV1 (small volume lcr reaction product: 4uL)
W3 BL05 LCR SV2
W4 BL05 LCR LV1 (large volume lcr reaction product: 16uL)
W5 BL05 LCR LV2
W6 BL01 LCR SV1
W7 BL01 LCR SV2
W8 Empty CMV/MV9
W9 BL01 LCR LV1
W10 BL01 LCR LV2
W11 LCR Negative Control Colony
W12 BL05 Traditional Ligation SV1
W13 BL05 Traditional Ligation SV2
W14 BL05 Traditional Ligation LV1
Ladder

DNA 4.0 μL
10X buffer 2.0 μL
XbaI 1.0 μL
NruI 1.0 μL
dH2O 12 μL
Total 20 μL --> 37°C/ 30 min.


Well 9 (BL01 LCR) looks promising! Will sequence after all samples are digested.