Description: The kit allows the comparative and quantitative detection of progerin expression by Real-Time PCR in cDNA samples and by Real-Time PCR, totally discriminating this specific target from the wild-type form, lamin A. The kit contents is sufficient for up to 40 determinations, including two calibration curves for lamin A and progerin (quantitative method) and two further amplification mixes for two reference genes necessary for the normalization of the expression data (comparative method).
Principal of the assay: The Progerin/Lamin A qPCR Detection Kit allows two possible methods of analysis: quantitative and comparative. Two positive standard controls for lamin A and progerin are included in the kit to perform the calibration curves needed to derive the copies number of the two target transcripts in the sample. The primers included in the optimized Lamin A Master Mix recognize a specific cDNA region that is present in the human lamin A coding sequence and absent in human progerin coding sequence. From this amplification reaction originates a PCR product of 179bp. Optimized Progerin Master Mix includes primers that recognize a specific human cDNA region flanking the deletion of 150 nucleotides that characterized the progerin transcript. From this amplification reaction originates a PCR product of 160 bp. Primers design have been accurately designed with the aim of maximizing the ability to discriminate progerin against wild-type lamin A.
Sensibility of the assay
- LOQ, Limit of Quantification: down to 100 lamin A and progerin copies
- LOD, Limit of Detection: down to 10 lamin A and progerin copies