Fluorescence level in real-time - (Apr/29/2008 )
Hi all,
I need urgent help on this topic.
1. How do I access the total fluorescence level emitted by the DNA product?
2. I'd constructed a standard curve of fluorescence level versus RNA concentration. I got a negative equation ie. as the RNA template concentration increases, the fluorescence level decreases. I'm doing one step RT-PCR. What are the possible explanation?
Help!!!!!
-audreyk-
QUOTE (audreyk @ Apr 29 2008, 11:23 AM)
Hi all,
I need urgent help on this topic.
1. How do I access the total fluorescence level emitted by the DNA product?
2. I'd constructed a standard curve of fluorescence level versus RNA concentration. I got a negative equation ie. as the RNA template concentration increases, the fluorescence level decreases. I'm doing one step RT-PCR. What are the possible explanation?
Help!!!!!
I need urgent help on this topic.
1. How do I access the total fluorescence level emitted by the DNA product?
2. I'd constructed a standard curve of fluorescence level versus RNA concentration. I got a negative equation ie. as the RNA template concentration increases, the fluorescence level decreases. I'm doing one step RT-PCR. What are the possible explanation?
Help!!!!!
You are correct, with the template concentration increasing, Ct value (reflected by fluorescence intensity) will decrease. The equation coefficient is negative.
-larryking-
Are you working on log concentrations?!
QUOTE (audreyk @ Apr 29 2008, 03:23 PM)
Hi all,
I need urgent help on this topic.
1. How do I access the total fluorescence level emitted by the DNA product?
2. I'd constructed a standard curve of fluorescence level versus RNA concentration. I got a negative equation ie. as the RNA template concentration increases, the fluorescence level decreases. I'm doing one step RT-PCR. What are the possible explanation?
Help!!!!!
I need urgent help on this topic.
1. How do I access the total fluorescence level emitted by the DNA product?
2. I'd constructed a standard curve of fluorescence level versus RNA concentration. I got a negative equation ie. as the RNA template concentration increases, the fluorescence level decreases. I'm doing one step RT-PCR. What are the possible explanation?
Help!!!!!
-kikinho-