it is not very different. just one more thing to add in your master mix per gene. I would just follow the manufacurer's insturctions.

What about to follow a protocol available on:

https://www.lifetechnologies.com/cz/en/home/life-science/pcr/real-time-pcr/real-time-pcr-assays/taqman-gene-expression/single-tube-taqman-gene-expression-analysis.html?ICID=search-product

I hope this helps.

Luděk

The main differences are :

- the use of a probe (for the quenching). You could use already made probes (commercial or from db) or define your own primers and probes using primer3.

- the enzyme/reaction mix, (not the mix of sybr green mix, no fluorescent molecule, not the same enzyme). I use an already made mix for TaqMan reaction.

- the wavelenght and the method of fluorescence detection (My qPCR system is a LC480).

Stephane

http://medgen.ugent.be/rtprimerdb/

Hi,

The Master Mix will be different from SYBR green master mix. There are Taqman Master mix available.

There will be no need to do Melting Curve step and analysis, as you will have specific PCR product.

You must know which fluorescence dyes are used in your Probes and even you can do Multi-plex PCR in one reaction, if the probes and fluorescence are different.

Good Luck.

hi

you can visit invitrogen web page :

http://www.lifetechnologies.com/us/en/home/brands/invitrogen.html

it is not very different. just one more thing to add in your master mix per gene. I would just follow the manufacurer's insturctions.

Taqman real time PCR is the quantitative PCR method which uses fluorogenic single stranded oligonucleotide probes. It binds to DNA sequence between the 2 PCR primers and generates fluorescent signals specific to the target. The protocol can be found on the website.and it is instrument specific.