Taqman versus SYBR Green - (May/05/2006 )
Hi!
I´m going to run a real-time PCR on my samples and my question is if I´m going to use taqman or SYBR Green.
I know that it is more specific to use a probe, but I intend to run the positive samples on a gel to confirm the size if I use SYBR Green. There are a lot of variants of the virus that I want to screen for so I want the PCR to pick up the virus even if they are not 100% identical, so maybe taqman is to specific?
Is any of the methods more sensitive than the other. I´ve read that SYBR Green has a sensitivity of 10 copies, but I have a test with Taqman that has a sensitivity of 1 copy. Is it just how you optimize the reaction?
I´m going to run a real-time PCR on my samples and my question is if I´m going to use taqman or SYBR Green.
I know that it is more specific to use a probe, but I intend to run the positive samples on a gel to confirm the size if I use SYBR Green. There are a lot of variants of the virus that I want to screen for so I want the PCR to pick up the virus even if they are not 100% identical, so maybe taqman is to specific?
Is any of the methods more sensitive than the other. I´ve read that SYBR Green has a sensitivity of 10 copies, but I have a test with Taqman that has a sensitivity of 1 copy. Is it just how you optimize the reaction?
Dear Johku,
What sort of virus are you sreening, DNA or RNA virus?
How many viruses are you going to screen? Are they in the same amily or different family?
TaqMan require high specificity, if there is some miss match bases at the probe region, your probe cant bind properly.
Since you are dealling with many viruses, i suggest you go with SYBR Green.
Best regards
Thank you for your answer!
I have now decided to use SYBR Green and I have reached the sensivity of one copy so that doesn't seem to be a difference between the two methods.