Endy:dNTP stock solution

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Current revision (10:58, 1 September 2005) (view source)
 
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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==
 
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#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).
 
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#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. <br>[[User:ClarkeS|Sean]] 17:50, 30 Aug 2005 (EDT)
 
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==Protocol #2==
 
My dNTP stocks are:
My dNTP stocks are:
*100x: 25mM each dNTP. Combine equal volumes of 100 mM individual dNTPs.
*100x: 25mM each dNTP. Combine equal volumes of 100 mM individual dNTPs.
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*Working concentration: 250 &mu;M<br>
*Working concentration: 250 &mu;M<br>
-[[User:Jkm|Jkm]] 10:04, 31 Aug 2005 (EDT)
-[[User:Jkm|Jkm]] 10:04, 31 Aug 2005 (EDT)
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This seems to make more sense than diluting individual dNTPs, the advantages being fewer tubes, no wondering if there's the same amount of each dNTP when you combine them, and fewer freeze-thaw cycles.
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[[User:ClarkeS|Sean]] 11:58, 1 Sep 2005 (EDT)

Current revision

Each individual dNTP comes as 25 μmol in 100 mM solution. The lids are color-coded to each nucleic acid.

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)

This seems to make more sense than diluting individual dNTPs, the advantages being fewer tubes, no wondering if there's the same amount of each dNTP when you combine them, and fewer freeze-thaw cycles. Sean 11:58, 1 Sep 2005 (EDT)

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