User:Karmella Haynes/Notebook/BioBrick cloning/2013/09/09: Difference between revisions

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'''DNA Concentration Data'''
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<!-- Below that, each | starts a row of cells. -->
<!-- The symbol || separates cells in a row. Replace the --- in each cell with your data. -->
<!-- If you have only one plasmid, delete the |- under Plasmid 1's row, and delete the entire row for Plasmid 2.-->
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{| {{table}}
|- bgcolor=#cfcfcf
| Plasmid || OD260 || OD260/280 || ng/μL
|-
| 1. Plasmid 1 || 0.609 || 1.85 || 609.4
|-
| 2. Plasmid 2 || 0.272 || 1.88 || 272.3
|}
'''Restriction Digest Table'''<br>
* Checked plasmid minipreps with EcoRI/PstI digests
{| {{table}} border="1" cellspacing="3"
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<!-- bgcolor=#cfcfcf *colors* the *background* of the Reagent and Volume cells grey.  -->
<!-- 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. -->
<!-- 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
<!-- After you upload your gel image to OWW, replace "GelImage.jpg" with the name of your image file -->
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|- valign="top"
| bgcolor=#cfcfcf | Reagent
| bgcolor=#cfcfcf | Volume
| rowspan="7" | <u>Expected:</u><br>1. CMV/V0120 colony 1 = 588, 3200<br>2. CMV/V0120 colony 2 = 588, 3200<br>3. CMV/V0120 (control) = 588, 3200<br>
| rowspan="7" | [[Image:Session3_JDS_digest.jpg‎|200px|Today's gel]]<br>[[Image:Session3_HB_digest.jpg‎|200px|Today's gel]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
|-
| DNA(plasmid) || 3.0 μL
|-
| 10X buffer || 1.5
|-
| EcoRI || 1.0
|-
| PstI || 1.0
|-
| dH<sub>2</sub>O || 8.5
|- valign="top"
| &nbsp; || 15 μL --> 37°C/ 15 min.
|}
'''DNA Sequencing Samples'''
<!-- * signs create an automatically bulleted list. Replace 'date' with the date you submitted the samples -->
* Submitted to DNASU on: '09/09/13'
* Order number: 8011
<!-- # 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. -->
# CMV/V0120 (control) - forward primer 'P0001'
# CMV/V0120 (control) - reverse primer 'P0002'
Note: Submitted along with Rene's reactions. Mine are samples 15 and 16 in the sequencing order.
* DNASU sequencing core power failure on 9/10/13 - data was lost
* Resubmitted samples on 9/11/13 as order 8018




Revision as of 17:49, 12 September 2013

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09/09/13

  • CMV/V0120 - minipreps, restriction digest, sequencing


  • CMV/V0120
    • Cultures were inoculated on 9/7/13
    • Culture 1 - growth, culture 2 - growth
    • Streak 1 - growth, streak 2 growth


Samples for Miniprep and Analysis

  1. Sample 1 = CMV/V0120 colony 1; BBa_J176027; part = 588 bp; vector = 3200 bp
  2. Sample 2 = CMV/V0120 colony 2; same
  3. Sample 3 = CMV/V0120 (control DNA); BBa_J176027; part = 588 bp; vector = 3200 bp


DNA Concentration Data

Plasmid OD260 OD260/280 ng/μL
1. Plasmid 1 0.609 1.85 609.4
2. Plasmid 2 0.272 1.88 272.3


Restriction Digest Table

  • Checked plasmid minipreps with EcoRI/PstI digests
Reagent Volume Expected:
1. CMV/V0120 colony 1 = 588, 3200
2. CMV/V0120 colony 2 = 588, 3200
3. CMV/V0120 (control) = 588, 3200
 Today's gel
Today's gel
15 μL/lane; 1% agarose; Ladder
DNA(plasmid) 3.0 μL
10X buffer 1.5
EcoRI 1.0
PstI 1.0
dH2O 8.5
  15 μL --> 37°C/ 15 min.


DNA Sequencing Samples

  • Submitted to DNASU on: '09/09/13'
  • Order number: 8011
  1. CMV/V0120 (control) - forward primer 'P0001'
  2. CMV/V0120 (control) - reverse primer 'P0002'

Note: Submitted along with Rene's reactions. Mine are samples 15 and 16 in the sequencing order.

  • DNASU sequencing core power failure on 9/10/13 - data was lost
  • Resubmitted samples on 9/11/13 as order 8018



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