Difference between revisions of "Haynes Lab:Notebook/David Barclay Undergrad Training/Plasmid Transformation/2013/10/03"

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(Continuation of Restriction Digests and Gel Electrophoresis)
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| bgcolor=#cfcfcf | Volume
 
| bgcolor=#cfcfcf | Volume
 
| rowspan="7" | <u>Expected:</u><br>1. fshPCD = 186 bp, 4968 bp<br>
 
| rowspan="7" | <u>Expected:</u><br>1. fshPCD = 186 bp, 4968 bp<br>
| rowspan="7" | [[Image:IMAG0329.jpg|400px|Today's gel]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
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| rowspan="7" | [[Image:Gel DKB 10-3-13.jpg|400px|Today's gel]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
 
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| DNA(plasmid) || 5.0 μL
 
| DNA(plasmid) || 5.0 μL
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'''Conclusion'''
 
'''Conclusion'''
 
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There were faint bands at the 186 position; however, they are harder to see on the picture. The 4968 bands for all 3 are very visible. Fairly successful digest.
  
 
'''DNA Sequencing Samples'''
 
'''DNA Sequencing Samples'''

Revision as of 22:21, 3 October 2013

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Continuation of Restriction Digests and Gel Electrophoresis

Restriction Digest Table

  • Checked plasmid minipreps with EcoRI/PstI digests
Reagent Volume Expected:
1. fshPCD = 186 bp, 4968 bp
Today's gel
15 μL/lane; 1% agarose; Ladder
DNA(plasmid) 5.0 μL
10X FD Buffer 1.5
EcoRI 1.0
PstI 1.0
dH2O 6.5
  15 μL --> 37°C/ 15 min.

Conclusion There were faint bands at the 186 position; however, they are harder to see on the picture. The 4968 bands for all 3 are very visible. Fairly successful digest.

DNA Sequencing Samples

  • Submitted to DNASU on:
  • Order Number:
  1. Plasmid 1 (fshPCD) - Forward P001
  2. Plasmid 1 (fshPCD) - Reverse P002