Endy:dNTP stock solution: Difference between revisions
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Each individual dNTP comes as 25 μmol in 100 mM solution. The lids are color-coded to each nucleic acid. | Each individual dNTP comes as 25 μmol in 100 mM solution. The lids are color-coded to each nucleic acid. | ||
==Protocol #1== | |||
#Dilute these in DI water to 2.5 mM to get four tubes of diluted dNTP, i.e., n*(1 μl dNTP + 39 μl water). | #Dilute these in DI water to 2.5 mM to get four tubes of diluted dNTP, i.e., n*(1 μl dNTP + 39 μl water). | ||
#Combine these in equal amounts to yield a 10x solution for use in PCR. | #Combine these in equal amounts to yield a 10x solution for use in PCR. | ||
I don't know if there's any advantage other than to minimize freeze-thaw cycles to keeping intermediate dilutions of individual dNTPs rather than combining them immediately into the 10x solution. [[User:ClarkeS|Sean]] 17:50, 30 Aug 2005 (EDT) | I don't know if there's any advantage other than to minimize freeze-thaw cycles to keeping intermediate dilutions of individual dNTPs rather than combining them immediately into the 10x solution. <br>[[User:ClarkeS|Sean]] 17:50, 30 Aug 2005 (EDT) | ||
==Protocol #2== | |||
My dNTP stocks are: | My dNTP stocks are: | ||
*100x: 25mM each dNTP. Combine equal volumes | *100x: 25mM each dNTP. Combine equal volumes of 100 mM individual dNTPs. | ||
*10x: 2.5mM each dNTP. Dilute back 100x stock 1:10. | *10x: 2.5mM each dNTP. Dilute back 100x stock 1:10. | ||
Working concentration: 250 μM<br> | *Working concentration: 250 μM<br> | ||
-[[User:Jkm|Jkm]] 10:04, 31 Aug 2005 (EDT) | -[[User:Jkm|Jkm]] 10:04, 31 Aug 2005 (EDT) | ||
Revision as of 11:52, 31 August 2005
Each individual dNTP comes as 25 μmol in 100 mM solution. The lids are color-coded to each nucleic acid.
Protocol #1
- Dilute these in DI water to 2.5 mM to get four tubes of diluted dNTP, i.e., n*(1 μl dNTP + 39 μl water).
- Combine these in equal amounts to yield a 10x solution for use in PCR.
I don't know if there's any advantage other than to minimize freeze-thaw cycles to keeping intermediate dilutions of individual dNTPs rather than combining them immediately into the 10x solution.
Sean 17:50, 30 Aug 2005 (EDT)
Protocol #2
My dNTP stocks are:
- 100x: 25mM each dNTP. Combine equal volumes of 100 mM individual dNTPs.
- 10x: 2.5mM each dNTP. Dilute back 100x stock 1:10.
- Working concentration: 250 μM
-Jkm 10:04, 31 Aug 2005 (EDT)
