Method and kit for the rapid sequencing of the human ATM gene
Rapid process for simultaneous sequencing of all gene exons in order to find any mutations for confirmation of AT (Atassia Telangiectasia) clinical diagnosis and for carriers.
The kit consists of 65 pairs of primers necessary for the amplification of the entire gene which are distributed in a multi-well plate, ready to undergo a single simultaneous amplification procedure (PCR). The procedure requires that the subsequent purification and sequence reaction steps are always carried out on a plate, using a multichannel pipette. The distribution of primers by sequence also follows a specific scheme so as not to alter the order in which the amplified exons are arranged. It is therefore possible to carry out the sequencing of the entire ATM gene with a few and simultaneous steps in order to significantly reduce the time (time required for the execution of the entire protocol is 2 working days, compared to the current 20) and costs (economic savings of about € 70 compared to to the traditional technique, all volumes and concentrations are calculated to measure, without waste but optimized to guarantee the final result). Compared to the current Sanger sequencing method, the different steps for sample preparation are performed simultaneously for all exons in the gene rather than a limited number of exons at a time. The entire procedure was tested on different DNA samples already characterized to verify the results. Perfect match of results.
- Molecular diagnostics laboratories;
- Research laboratories.
- Rapid sequencing;
- Simultaneous preparation of all exons of the gene to be analyzed;
- Reduces the possibility of error;
- Much cheaper system than the single preparation of each exon to be sequenced;
- It does not require additional techniques to be validated unlike the NGS.