Haynes Lab:Notebook/Short Projects/2014/09/18: Difference between revisions

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Haynes Lab - Short Projects</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Haynes Lab - Short Projects</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
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'''Plasmid sequencing - planning'''
'''Plasmid sequencing - planning'''
* Prep primers
# P0001 (blue cap = 100 μM)
# P0002 (blue cap = 100 μM)
* Thaw stock primers (blue capped tubes) at room temp.
* Get two sterile 1.5 mL tubes
* Label each with the primer name and "10 μM"
* Add 90 μL dH<sub>2</sub>O to each tube
* Transfer 10 μL P0001 from blue caped tube into 90 μL H<sub>2</sub>O in new tube. Final volume = 100 μL, final conce. = 10 μM
* Repeat last step for P0002.
* Use DNASU: http://openwetware.org/wiki/Haynes:TechResources#Ordering
* Use DNASU: http://openwetware.org/wiki/Haynes:TechResources#Ordering


{| {{table}}
{| {{table}}
|-
|-
| Tube || Primer (1 μL of 10 μM) || DNA (200 ng)
| Tube || Primer (1 μL of 10 μM) || DNA (200 ng) || [DNA] || '''Vol DNA''' || '''Vol 10 μM primer''' || '''Vol dH<sub>2</sub>O'''
|-
|-
| 1. || P0001 || BD003
| 1. || P0001 || BD003 || 128 ng/μL || 1.6 μL || 1.0 μL || 7.4 μL
|-
|-
| 2. || P0002 || BD003
| 2. || P0002 || BD003 || 128 ng/μL || 1.6 μL || 1.0 μL || 7.4 μL
|-
|-
| 3. || P0001 || BD004
| 3. || P0001 || BD004 || 141 ng/μL || 1.4 μL || 1.0 μL || 7.6 μL
|-
|-
| 4. || P0002 || BD004
| 4. || P0002 || BD004 || 141 ng/μL || 1.4 μL || 1.0 μL || 7.6 μL
|-
|-
| 5. || P0001 || BD005
| 5. || P0001 || BD005 || 290.2 ng/μL || 0.7 μL || 1.0 μL || 8.3 μL
|-
|-
| 6. || P0002 || BD005
| 6. || P0002 || BD005 || 290.2 ng/μL || 0.7 μL || 1.0 μL || 8.3 μL
|-
|-
| 7. || P0001 || BD006
| 7. || P0001 || BD006 || 255.3 ng/μL || 0.8 μL || 1.0 μL || 8.2 μL
|-
|-
| 8. || P0002 || BD006
| 8. || P0002 || BD006 || 255.3 ng/μL || 0.8 μL || 1.0 μL || 8.2 μL
|}
|}


 
* Use 0.2 mL PCR strip tubes
* Be sure to leabel each one
* Place in a plastic baggie (recycled)
* Use tape to secure baggie with tubes to order form




Latest revision as of 00:19, 27 September 2017

Haynes Lab - Short Projects Main project page
Previous entry      Next entry

09/18/14 - Meli'sa Crawford

  • Plasmid sequencing - planning


Plasmid sequencing - planning

  • Prep primers
  1. P0001 (blue cap = 100 μM)
  2. P0002 (blue cap = 100 μM)
  • Thaw stock primers (blue capped tubes) at room temp.
  • Get two sterile 1.5 mL tubes
  • Label each with the primer name and "10 μM"
  • Add 90 μL dH2O to each tube
  • Transfer 10 μL P0001 from blue caped tube into 90 μL H2O in new tube. Final volume = 100 μL, final conce. = 10 μM
  • Repeat last step for P0002.


Tube Primer (1 μL of 10 μM) DNA (200 ng) [DNA] Vol DNA Vol 10 μM primer Vol dH2O
1. P0001 BD003 128 ng/μL 1.6 μL 1.0 μL 7.4 μL
2. P0002 BD003 128 ng/μL 1.6 μL 1.0 μL 7.4 μL
3. P0001 BD004 141 ng/μL 1.4 μL 1.0 μL 7.6 μL
4. P0002 BD004 141 ng/μL 1.4 μL 1.0 μL 7.6 μL
5. P0001 BD005 290.2 ng/μL 0.7 μL 1.0 μL 8.3 μL
6. P0002 BD005 290.2 ng/μL 0.7 μL 1.0 μL 8.3 μL
7. P0001 BD006 255.3 ng/μL 0.8 μL 1.0 μL 8.2 μL
8. P0002 BD006 255.3 ng/μL 0.8 μL 1.0 μL 8.2 μL
  • Use 0.2 mL PCR strip tubes
  • Be sure to leabel each one
  • Place in a plastic baggie (recycled)
  • Use tape to secure baggie with tubes to order form




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