Haynes Lab:Notebook/Engineering PC-TFs/2014/07/09: Difference between revisions

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==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="3"
|- valign="top"
| bgcolor=#cfcfcf | Reagent
| bgcolor=#cfcfcf | Volume
| rowspan="7" | <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="7" | [[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="7" | [[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.
Bottom W2: BL05 possible success.
Bottom W4: BL09 possible success.
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__NOTOC__
__NOTOC__

Revision as of 13:45, 10 July 2014



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Summary

Liquid Culture Minipreps
Eluted with 70μL dH2O.

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


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


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


Restriction Digest

Reagent Volume Expected:
BL01(W1:2-4), BL05(W1:5, W2:2-3)
Forward insertion: 710, 7010.
Reverse insertion: 3230, 4490.
No insertion: 710, 4490.
BL09 (W2:4-6)
Forward insertion: 710, 5921
Reverse insertion: 2141, 4490
No insertion: 710, 4490.
*Final well is empty CMV/MV9. 		Today's gel
15 μL/lane; 1% agarose;
Observed lengths: W2: 7100, 700; W3: 7100, 5200, 2500; W4: *5200, 2500, 710; W5: 5000, 2500; W6: 5000, 2500, 710.
*Uncut plasmid.
Ladder 		Today's gel
15 μL/lane; 1% agarose;
Observed lengths: W2: *7100, 700; W3: *6100, 2500, 710; W4: *7200, 2500, 710; W5: 5000, 4000; W6: *5500, 2500, 750.
*Uncut plasmid.
Ladder
DNA(BL01x3, BL05x3, BL09x3, CMV/MV9) 5.0 μL
10X buffer 1.5 μL
XbaI 1.0 μL
NruI 1.0 μL
dH2O 6.5 μL
Total 15 μL --> 37°C/ 30 min.


Results
Top W2: BL01 possible success. Bottom W2: BL05 possible success. Bottom W4: BL09 possible success.


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