qRT-PCR - data analysis - methods that DO NOT use a calibrator ! (Sep/29/2008 )
hi,
I have a question regarding qRT-PCR data analysis: which method should be used to quantify gene expression in different organs, normalized to an internal control BUT without using one of the organ sample as calibrator?
Thank you!
diana
-diana c-
hi diana,
what do you want to quantify in detail?
-> use a just the ratio between target and reference gene assay if this information is enough?!
->use a general calibrator like the mixture of your samples. But better to use a calibrator...
best wishes
the professor
QUOTE (diana c @ Sep 29 2008, 11:17 AM)
hi,
I have a question regarding qRT-PCR data analysis: which method should be used to quantify gene expression in different organs, normalized to an internal control BUT without using one of the organ sample as calibrator?
Thank you!
diana
I have a question regarding qRT-PCR data analysis: which method should be used to quantify gene expression in different organs, normalized to an internal control BUT without using one of the organ sample as calibrator?
Thank you!
diana
-THE_PROFESSOR-
QUOTE (THE_PROFESSOR @ Sep 29 2008, 10:18 AM)
hi diana,
what do you want to quantify in detail?
-> use a just the ratio between target and reference gene assay if this information is enough?!
->use a general calibrator like the mixture of your samples. But better to use a calibrator...
best wishes
the professor
what do you want to quantify in detail?
-> use a just the ratio between target and reference gene assay if this information is enough?!
->use a general calibrator like the mixture of your samples. But better to use a calibrator...
best wishes
the professor
QUOTE (diana c @ Sep 29 2008, 11:17 AM)
hi,
I have a question regarding qRT-PCR data analysis: which method should be used to quantify gene expression in different organs, normalized to an internal control BUT without using one of the organ sample as calibrator?
Thank you!
diana
I have a question regarding qRT-PCR data analysis: which method should be used to quantify gene expression in different organs, normalized to an internal control BUT without using one of the organ sample as calibrator?
Thank you!
diana
Thank you for your suggestions and I will try to see if they would fit to my experiment design!
I want to see the expression pattern of "n" genes in different Arabidopsis organs;thus, I;m using TaqMan and Act2 as reference, but since I don't have treated/untreated samples or wild type/mutant (samples are only from wild type plants),it doesn't make sense to choose one of the samples(e.g. one of the organs) as calibrator!
All the best!
Diana
-diana c-
you dont need an callibrator, just messure for each sample your GOI and your control gene substract them from each other and you have a way to compare your samples among each other.
-hebus-