The fluorescence curve-2

[Makky]

:doh:Hi guys, I'll use RT-PCR device to identify some bacterial species on the 16S rRND level using F and R primers. DNA was extracted from the bacterial samples and used as a template and purified DNA containing the same primer sequence will use as a reference for comparison the samples results and the reference result. The primer sheet reported that the Tm (melting temperature) of primer is 60.4C and the PCR protocol run using SYBR Green-1 using melting curve analysis.

I need to know exactly If my sample is amplified and was specific and closely related to the reference, Is the sample must give the same Tm that for primer sheet (60.4 C) from melting curve?? or How I Identify my bacterial samples?? OR record the new one for reference and compare it to the sample??

How I can measure the fluorescence curve obtained from my machine (Bio-Rad)?

 

Plz, I need breifly help in this point.

 

Many thanks,