Centogene
EXTAN PMP22 Kit
OMIM 601097
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Extan Kit for PMP22 by Centogene was used to identify point mutations responsible for CMT1A / CMT1E. The instructions in the kit for the setting up of experiment were explicit making the kit very easy to use. Sanger sequencing of the PCR products amplified using primer sets provided in the kit gave us satisfactory results. Dosage analysis of PMP22 gene which is already established in our laboratory along with the EXTAN PMP22 kit will help us to delineate the molecular defect responsible for causing Charcot–Marie–Tooth disease.
Diagnosis
DNA
The testing for PMP22 gene was done using EXTAN PMP22 kit provided by CENTOGENE. Protocol:The primers and Positive Amplification Control (PAC) were reconstituted according to the Manufacturer’s guidelines.We have used three different Taq polymerases to amplify the fragments. Taq polymerases used were: 1) Bangalore GeNei (Taq 3U/microL; 10X buffer)2) AmpliTaq Gold (Taq 5U/microL; MgCl2, 10X buffer II)3) HotStar Taq Plus Polymerase (Taq 5U/microL; 10X buffer; 5X Q Solution)PCR was set up for the three test DNA samples provided by the Manufacturer along with the PAC and negative control. PCR program used was “CENTO 1” with a modification in the initial denaturation step (Instead of 15 minutes, we used 5 minutes only as initial denaturation)
Stick to the Manufacturer's instructions for Best results
With HotStart Taq, we were able to amplify all specific fragments for all the three Test DNA samples along with the PAC. Sequencing PCR was setup using Big Dye Terminator v3.1 Cycle Sequencing kit. Sequencing results were analysed using SeqScape and Blated using UCSC Genome Browser. Overall, the results obtained are Satisfactory
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easy to use
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Easy to use product. No hassle to create primers and standardization.
Product Name: EXTAN® PMP22 Primer Set KitSupplier Page