Endy:dNTP stock solution

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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.
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==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.
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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)
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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)
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==Protocol #2==
My dNTP stocks are:
My dNTP stocks are:
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*100x: 25mM each dNTP. Combine equal volumes each individual dNTP.
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*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.
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Working concentration: 250 &mu;M<br>
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*Working concentration: 250 &mu;M<br>
-[[User:Jkm|Jkm]] 10:04, 31 Aug 2005 (EDT)
-[[User:Jkm|Jkm]] 10:04, 31 Aug 2005 (EDT)

Revision as of 14:52, 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

  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).
  2. 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)

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