Haynes Lab:Notebook/David Barclay Undergrad Training/Plasmid Transformation/2013/10/01: Difference between revisions

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=Day 3- Minipreps fshPCD=
==Liquid Culture Results==
NOTE: Cultures and Streaks taken out after 18 hours (37 incubator) and left on lab bench for 1 hour.
===fshPCD===
*Cultures Innoculated [http://openwetware.org/wiki/Haynes_Lab:Notebook/David_Barclay_Undergrad_Training/Plasmid_Transformation/2013/09/27 09/27/13]
*Cultures 1 and 2: Both show growth
*Streak 1 and 2: Both show growth


==Samples for Minipreps and Analysis==
*Sample 1= fshPCD/V0120 colony 1; BBa_J176002; part= 186 bp; vector= 4968 bp
*Sample 2= hPCD/V0120 colony 2; same
=='''DNA Concentration Data'''==
<!-- Wiki code notes: The {| is the opening tag for tables. {{table}} is an OWW template that creates thin grey border lines and cell margins.
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<!-- The symbol || separates cells in a row. Replace the --- in each cell with your data. -->
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{| {{table}}
|- bgcolor=#cfcfcf
| Plasmid || OD260 || OD260/280 || ng/μL
|-
| 1. fshPCD 1|| 0.014 || 1.714 || 24.682|-
| 2. fshPCD 2 || 0.013 || 1.762 || 28.523|}
'''Did not have good concentration, nothing else done after this point'''
Restriction Digest Table<br>
* Checked plasmid minipreps with EcoRI/PstI digests
{| {{table}} border="1" cellspacing="3"
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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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|- valign="top"
| bgcolor=#cfcfcf | Reagent
| bgcolor=#cfcfcf | Volume
| rowspan="7" | <u>Expected:</u><br>1. bp<br>2. bp<br>
| rowspan="7" | [[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 FD Buffer || 1.5
|-
| EcoRI || 1.0
|-
| PstI || 1.0
|-
| dH<sub>2</sub>O || 8.5
|-
| &nbsp; || 15 μL --> 37°C/ 15 min.
|}
'''Conclusion'''
'''DNA Sequencing Samples'''
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<!-- The Order Number is shown at the top of your sequencing order form. Record in the indicated spot below -->
* Submitted to DNASU on:
* Order Number:
<!-- # 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. -->
# Plasmid 1
# Plasmid 2




Revision as of 08:38, 2 October 2013

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Day 3- Minipreps fshPCD

Liquid Culture Results

NOTE: Cultures and Streaks taken out after 18 hours (37 incubator) and left on lab bench for 1 hour.

fshPCD

  • Cultures Innoculated 09/27/13
  • Cultures 1 and 2: Both show growth
  • Streak 1 and 2: Both show growth

Samples for Minipreps and Analysis

  • Sample 1= fshPCD/V0120 colony 1; BBa_J176002; part= 186 bp; vector= 4968 bp
  • Sample 2= hPCD/V0120 colony 2; same

DNA Concentration Data

Plasmid OD260 OD260/280 ng/μL
1. fshPCD 1 0.014 1.714 - 2. fshPCD 2 0.013 1.762 }

Did not have good concentration, nothing else done after this point

Restriction Digest Table

  • Checked plasmid minipreps with EcoRI/PstI digests
Reagent Volume Expected:
1. bp
2. bp
 Today's gel
15 μL/lane; 1% agarose; Ladder
DNA(plasmid) 3.0 μL
10X FD Buffer 1.5
EcoRI 1.0
PstI 1.0
dH2O 8.5
  15 μL --> 37°C/ 15 min.

Conclusion


DNA Sequencing Samples

  • Submitted to DNASU on:
  • Order Number:
  1. Plasmid 1
  2. Plasmid 2
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