Thus while a small experiment using a low starting quantity of RNA may fail to identify many subtle changes, one can have confidence in any changes that are reported.
Therefore, a higher Ct value indicates a lower starting quantity of nucleic acid.
Even with the advent of whole genome amplification, to achieve representative amplification of the genome a minimum starting quantity of 1 μl of 10 ng/μL is recommended (GE HealthCare GenomiPhi™™ instruction manual).
The median ratios of the two signal to noise ratios are 1.12, 1.16 and 1.76 for 100 ng, 50 ng or 10 ng respectively comparing to a reference starting quantity of 250 ng.
The higher the starting quantity of a target sequence, the earlier a significant increase in fluorescence is detected.
To determine RT PCR efficiency and initial starting quantity of the samples, a standard curve was generated using samples from a 1 : 3 dilution series of total starting cDNA.
The relative starting quantity of cDNA for a particular gene was determined in GENEX (Biorad) using the following equation: relative quantity = efficiency (control Ct-experimental Ct) based on Livak [ 129] and Vandesompele [ 130].
We find that the starting quantity of RNA has an effect on observed intensities despite the fact that the quantity of cRNA being hybridized remains constant.
The higher the starting quantity of the template, the earlier a significant increase in fluorescence is observed (Wittwer et al, 1997; Bieche et al, 1999).
The higher the starting quantity of the template, the earlier a significant increase in fluorescence is observed (Bieche et al, 1999).
