EXTAN PMP22 Primer Set

June 24, 2014

Overall

Quality of Results

Ease-of-Optimization

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National Genetic Center, Royal Hospital
Molecular Genetics Section
Clinical Scientist & Laboratory Technical Supervisor

Company:

CENTOGENE

Product Name:

EXTAN Primer set

Catalog Number:

S0001

EXTAN is a highly useful concept for diagnostic labs with limited resources and staff as it eliminates the need for designing molecular genetic tests from scratch. Diagnostic labs where many different genetic tests are carried out at the same time, will probably prefer EXTAN kits as the entire methodology is well explained from PCR to sequencing clean-up and require no further optimizations. The EXTAN set included excellently optimized primers with no primer dimers detected!

Experimental Design and Results Summary

Application

PMP22 (OMIM 601097) full gene sequencing

Starting Material

DNA

Protocol Overview

1) PCR carried on ABI Veriti Thermal Cycler exactly as described in the EXTAN Handbook. 2) PCR products were analyzed using Manual Gel Electrophoresis 3) PCR purification was carried out with:• Shrimp Alkaline Phosphatase (SAP); Cat. No. 70092Y/Z/X• Fermatas Exonuclease I (Exo I); Cat. No. EN05814) Cycle-sequencing was carried exactly as described in the EXTAN Handbook. 5) Sequencing was carried out on ABI 3130 (4-capillary) Sequencer. 6) Sequence data was compiled assembled and analyzed using ABI SeqScape software

Tips

There is little room for error due to the restriction of reagent quantities provided. This may pose a problem for first-time users, especially if funding is limited for tests.

Results Summary

Variants in the relevant exons and splice-sites were detected in three separate DNA samples. Known variants in a control DNA sample provided within the kit were also detected accurately.

Additional Notes

Only micromutations can be detected using the EXTAN Kits. Large duplications and deletions cannot be detected using this kit.

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Summary

The Good

Easy to follow work-flow. Great results using minimal reagent quantities.

The Bad

None

The Bottom Line

The EXTAN PMP22 test kit is an effective and convenient system for the analysis of micromutations within the clinically relevant regions of the PMP22 gene.

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