Lidstrom:Diluting Primers: Difference between revisions
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(New page: Back to: Lidstrom Protocols Primer Dilutions: People generally dilute dehydrated primers that arrive in the mail to 100uM, and use a working concentration of 10 uM....) |
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Primer Dilutions: | Primer Dilutions: | ||
People generally dilute dehydrated primers that arrive in the mail to 100uM, and use a working concentration of 10 uM. | People generally dilute dehydrated primers that arrive in the mail to 100uM, and use a working concentration of 10 uM. | ||
== Prepare 100 uM stock == | |||
*To get 100uM: dissolve dehydrated primer with in 10 uL per nmole: | |||
**Example: for 25 nmoles of primer add 250 ul of water | |||
***This will give you a concentration of 0.0001 M or 0.1 mM | |||
****(1 nmole/10uL) = (1 nmole/10uL)*(10^6 uL/L)*(10^-9 mole/nmole) = 0.0001M = 0.1 mM = 100 uM | |||
==Prepare primer for use== | |||
*Dilute this stock 1:10, to give a concentration of 10 uM | |||
==Use in PCR== | |||
*From this, use __ ml in a typical __ uL PCR reaction. This will give you a final concentration of __ pmoles in a PCR reaction. | |||
From this, use __ ml in a typical __ uL PCR reaction. This will give you a final concentration of __ pmoles in a PCR reaction. |
Revision as of 19:10, 18 January 2012
Back to: Lidstrom Protocols
Primer Dilutions: People generally dilute dehydrated primers that arrive in the mail to 100uM, and use a working concentration of 10 uM.
Prepare 100 uM stock
- To get 100uM: dissolve dehydrated primer with in 10 uL per nmole:
- Example: for 25 nmoles of primer add 250 ul of water
- This will give you a concentration of 0.0001 M or 0.1 mM
- (1 nmole/10uL) = (1 nmole/10uL)*(10^6 uL/L)*(10^-9 mole/nmole) = 0.0001M = 0.1 mM = 100 uM
- This will give you a concentration of 0.0001 M or 0.1 mM
- Example: for 25 nmoles of primer add 250 ul of water
Prepare primer for use
- Dilute this stock 1:10, to give a concentration of 10 uM
Use in PCR
- From this, use __ ml in a typical __ uL PCR reaction. This will give you a final concentration of __ pmoles in a PCR reaction.