Haynes Lab:Notebook/Engineering PC-TFs/2014/08/08

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(Autocreate 2014/08/08 Entry for Haynes_Lab:Notebook/Engineering_PC-TFs)
Current revision (17:26, 8 August 2014) (view source)
(Summary)
 
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==Summary==
==Summary==
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*  
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*Concentration reading for gel purified PcTFs, ligation product minipreps.
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*Gel verification of ligation attempt 3.
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----
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'''Concentration Readings'''
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<br>
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{| {{table}} border="1" cellspacing="3" <!-- Plates. table -->
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|- valign="top"
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| rowspan="7" | [[Image:Bl01,5,9 gel p conc.jpg|400px|Hover name]]
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|}
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<br>
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----
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'''Gel Verification'''
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{| {{table}} border="1" cellspacing="5"
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|- valign="top"
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| bgcolor=#cfcfcf | Reagent
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| bgcolor=#cfcfcf | Volume
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| rowspan="9" |
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<u><b>Expected:</b></u>
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<br>
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<br>BL01(Well: 2), BL05(Well:3-6) <br>
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Forward insertion: 710, 7010. <br>
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Reverse insertion: 3230, 4490. <br>
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No insertion: 710, 4490. <br><br>
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 +
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<u>CMV/MV9 [XbaI/NruI] (Well: 7)</u> <br>
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710, 4490<br><br>
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<u>BL09 Ligation Colonies 1-6[XbaI/NruI] (Well: 8-13)</u><br>
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Forward insertion: 710, 5921 <br>
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Reverse insertion: 2141, 4490 <br>
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No insertion: 710, 4490<br> <br>
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| rowspan="10" | [[Image:08 08 14 pctf lig3 ver20140808 141240.jpg|400px|Today's gel]]<br>
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15 μL/lane; 1% agarose;<br>
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<u>Observed lengths:</u> <br>
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W2(BL01Lig1): 5000, 700; <br>
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W3(BL05Lig1): 5000, 700; <br>
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W4(BL05Lig2): 5000, 700; <br>
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W5(BL05Lig3): 5000, 700; <br>
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W6(BL05Lig4): 5000, 700; <br>
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W7(Empty CMV/MV9): 5000, 700; <br>
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W8(BL09Lig1): 5000, 700; <br>
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W9(BL09Lig2): 7000, 700; <br>
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W10(BL09Lig3): 5000, 700; <br>
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W11(BL09Lig4): 5000, 700; <br>
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W12(BL09Lig5): 7000, 700;<br>
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W13(BL09Lig6): 5000, 700; <br>
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[http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
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|-
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| DNA(BL01x2, BL05x1, BL09x2, CMV/MV9x1) || 2.0 μL
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|-
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| 10X buffer || 1.5 μL
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|-
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| XbaI || 1.0 μL
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|-
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| NruI || 1.0 μL
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|-
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| dH<sub>2</sub>O || 9.5 μL
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|-
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| Total || 15 μL --> 37°C/ 30 min.
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|}
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<br>
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BL09Lig2&5 sent in for sequencing.
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Summary

  • Concentration reading for gel purified PcTFs, ligation product minipreps.
  • Gel verification of ligation attempt 3.

Concentration Readings

Hover name



Gel Verification

Reagent Volume

Expected:

BL01(Well: 2), BL05(Well:3-6)
Forward insertion: 710, 7010.
Reverse insertion: 3230, 4490.
No insertion: 710, 4490.


CMV/MV9 [XbaI/NruI] (Well: 7)
710, 4490


BL09 Ligation Colonies 1-6[XbaI/NruI] (Well: 8-13)
Forward insertion: 710, 5921
Reverse insertion: 2141, 4490
No insertion: 710, 4490

Today's gel

15 μL/lane; 1% agarose;

Observed lengths:
W2(BL01Lig1): 5000, 700;
W3(BL05Lig1): 5000, 700;
W4(BL05Lig2): 5000, 700;
W5(BL05Lig3): 5000, 700;
W6(BL05Lig4): 5000, 700;
W7(Empty CMV/MV9): 5000, 700;
W8(BL09Lig1): 5000, 700;
W9(BL09Lig2): 7000, 700;
W10(BL09Lig3): 5000, 700;
W11(BL09Lig4): 5000, 700;
W12(BL09Lig5): 7000, 700;
W13(BL09Lig6): 5000, 700;
Ladder

DNA(BL01x2, BL05x1, BL09x2, CMV/MV9x1) 2.0 μL
10X buffer 1.5 μL
XbaI 1.0 μL
NruI 1.0 μL
dH2O 9.5 μL
Total 15 μL --> 37°C/ 30 min.


BL09Lig2&5 sent in for sequencing.


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