Haynes Lab:Notebook/Short Projects/2014/09/18: Difference between revisions
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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 | |||
Revision as of 11:53, 19 September 2014
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09/18/14 - Meli'sa Crawford
Plasmid sequencing - planning
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