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Need help with diluting heterozygoud DNA

Need help with diluting heterozygoud DNA - DNA Forum

Need help with diluting heterozygoud DNA - Discuss DNA, the molecule of hereditary. Topics include DNA structure, DNA replication, DNA function.


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Old 10-07-2011, 12:24 PM
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Default Need help with diluting heterozygoud DNA



I am trying to make a series of sensitivity controls for a real time PCR assay I am setting up. I usually can do dilutions using 100% WT and 100% mutant cell line DNA. However this time, the cell line I grew was about 50% mutant at my hotspot. So lets say I have a 50% mutant (positive) cell line to start with and I use 100% normal genomic DNA for the diluent.

What do I pipette to get about 100ul of the following concentrations

50% mutant I can use straight
10% mutant =
1% mutant =
0.1% mutant =
0.01% mutant

I originally just did a 1 to 4 dilution on the straight pos cell line but realized that is not a 10% b/c itís a heterozygous allele.
Thanks for any help, much is needed.

kmt
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