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Hi,I am a little bit confused about the primer dilutions. It...

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Dani Wicaksono Nov 22, 2016
protocol Protocol: Telomerase Repeated Amplification Protocol (TRAP)
Hi,

I am a little bit confused about the primer dilutions. It was shown in the protocol that ACX and NT primers stock concentration are both 1 ug/ul respectively. Do I need to dissolve the primers to 1 ug/ul for the stocks? The "final concentration" of ACX and NT primers is 100 Read more Read less more less
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Jerry W. Shay Author Answered Nov 23, 2016

Department of Cell Biology, University of Texas Southwestern Medical Center, USA

the final primer concentration in the TRAP reaction is 2ng/uL.

The final primer concentrations are 100ng/ul in the primer mix. As listed in the table under 'recipes' the stock is 1ug/uL and the final in the primer mix is 100ng/uL. When 1ul of the primer mix is added to a 49 uL reaction (final volume of 50ul thus 100ng/50uL = 2ng/uL final primer concentration) the final concentration of primers is 2ng/uL in the trap extension/PCR.

The primers are used at lower concentration than typical PCR because of potential primer artifacts.
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