Taqman SNP assay

Duncan Clark blackhole at abuse.plus.com
Mon Oct 4 07:29:45 EST 2004


Historians believe that in newspost <41612A89.7030906 at uta.fi> on Mon, 4 
Oct 2004, Nina Mononen <nina.mononen at uta.fi> penned the following 
literary masterpiece:
>Assay mix is the one that has PCR primers and probes, master mix 
>contains enzymes etc (at least according to the instruction leaflets 
>that come with the assays : )
>

OK so all you have done is halved the primer/probe concentration.

Not really a problem and is what you often do with primer concentration 
in a SYBR reaction anyway (to reduce dimers).

We normally run 300nM primers and 100nM probe but you can alter that 
quite a lot without causing the result to fail.

Duncan

>Nina
>
>Duncan Clark wrote:
>
>> Historians believe that in newspost <4160F27F.2050704 at uta.fi> on Mon, 
>>4 Oct 2004, Nina Mononen <nina.mononen at uta.fi> penned the following 
>>literary masterpiece:
>>
>>> While working with my first TaqMan assay I accidentally added 0.625 
>>>ul of the assay mix to each reaction (instead of the 1.25 ul) and the 
>>>assay worked fine : ) So, was I just lucky? Do these assays generally 
>>>work with less amount of assay mix added than what is recommended on 
>>>the Applied Biosystems instructions?
>>
>>
>> You need to define what you mean by assay mix?
>>
>> You can't be meaning, with that volume, the mastermix so I'm guessing 
>>you mean amount of primer probe mix?
>>
>> Duncan
>
>

-- 
I love deadlines. I especially like the whooshing noise they make as
they go flying by.

Duncan Clark
GeneSys Ltd.



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