Haynes Lab:Notebook/David Barclay Undergrad Training/Plasmid Transformation/2013/10/03

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(Autocreate 2013/10/03 Entry for Haynes_Lab:Notebook/David_Barclay_Undergrad_Training/Plasmid_Transformation)
Current revision (16:17, 7 October 2013) (view source)
(Continuation of Restriction Digests and Gel Electrophoresis)
 
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=Continuation of Restriction Digests and Gel Electrophoresis=
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Restriction Digest Table<br>
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* Checked plasmid minipreps with EcoRI/PstI digests
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{| {{table}} border="1" cellspacing="3"
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<!-- Editing: the coding for this table is a bit more advanced. -->
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<!-- The next two "rowspan=7" cells span  all 7 rows in the table so that they can fit the "Expected" list and a gel image. rowspan is how you create merged rows in Wiki code. -->
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<!-- Replace Plasmid 1 and 2 with the names of your plasmids. Replace insert size and vector size with appropriate information for your plasmids. If you only have one Plasmid, delete all the text for Plasmid 2
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<!-- After you upload your gel image to OWW, replace "GelImage.jpg" with the name of your image file -->
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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="7" | <u>Expected:</u><br>1. fshPCD = 186 bp, 4968 bp<br>
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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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|-
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| DNA(plasmid) || 5.0 μL
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|-
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| 10X FD Buffer || 1.5
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|-
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| EcoRI || 1.0
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|-
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| PstI || 1.0
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|-
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| dH<sub>2</sub>O || 6.5
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|-
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| &nbsp; || 15 μL --> 37°C/ 15 min.
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|}
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'''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.
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'''DNA Sequencing Samples'''
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* Submitted to DNASU on: 10/7/2013
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* Order Number: 8213
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<!-- # signs create an automatically numbered list. Replace Plasmid 1 & 2 with the names of your plasmids. Replace 'name' with the appropriate name of each primer. -->
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# Plasmid 1 (fshPCD) - Forward P001
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# Plasmid 1 (fshPCD) - Reverse P002

Current revision

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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: 10/7/2013
  • Order Number: 8213
  1. Plasmid 1 (fshPCD) - Forward P001
  2. Plasmid 1 (fshPCD) - Reverse P002



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