Google Sign in options
Remember me
This is not recommended for shared computers

Sign in anonymously
Don't add me to the active users list

Privacy Policy
Sign In
Create Account

Javascript Disabled Detected

You currently have javascript disabled. Several functions may not work. Please re-enable javascript to access full functionality.

Select the optimal primer-concentration

Started by whitelines, Feb 15 2013 05:36 AM

real-time pcr primers concentration optimization

Please log in to reply

No replies to this topic

#1 whitelines

member

Members
1 posts

2
Neutral

Posted 15 February 2013 - 05:36 AM

Hi,

How do I decide the optimal primer-concentration for my assay? Available litterature says run a matrix of concentrations and choose the one with lowest Ct-value.

However I am concerned that the more substantial formation of primer-dimers at the concentration giving the lowest Ct (red arrows in figure) might have an impact on the sensitivity of the assay when working with lower concentrations of template?
(My goal is to procude the most sensitive assay possible (viral DNA). Primers are published and used by others.)

Figure:
Primer koncentration = 300nM (red arrows)
Primer concentration = 100nM (blue arrows)

Hope I could have som feedback on this one

Thanks in advance.

Back to PCR, RT-PCR and Real-Time PCR · Next Unread Topic →

Also tagged with one or more of these keywords: real-time pcr, primers, concentration, optimization

Protocols and Techniques Forums → PCR, RT-PCR and Real-Time PCR → using cell line to generate standard curve Started by Guest_detriar_* , 14 Apr 2013 cell line, standard curve and 1 more...	6 replies 261 views	detriar Today, 08:39 PM
Protocols and Techniques Forums → Microbiology → How to determine concentration of phage-like particles? Started by Guest_Guf_* , 25 Mar 2013 MS2 bacteriophage and 2 more...	0 replies 193 views	Guf 25 Mar 2013
Protocols and Techniques Forums → Molecular Biology → ligation with one ristrection site/enzyme Started by Guest_yusii_* , 16 Mar 2013 ristriction digest, ligation and 2 more...	3 replies 231 views	2xzwei 20 Mar 2013
Protocols and Techniques Forums → Protein and Proteomics → Determining protein concentration after solubilisation in Triton X-100 Started by Guest_Joe101_* , 21 Feb 2013 bradford, protein, concentration and 2 more...	2 replies 252 views	sam_neuwied 21 Feb 2013
Products and Vendors Discussion → Lab Equipments → How useful are Service Contracts for qPCR machines? Started by Guest_doxorubicin_* , 19 Feb 2013 qpcr, real-time pcr	2 replies 119 views	Tabaluga 19 Feb 2013