Sensitive Detection of Molecular Targets in Cancer by Minisequencing

Şirin Yüksel Kılıçturgay; Cemaliye Boylu Akyerli

doi:10.31067/acusaglik.851006

TR EN

Mini-dizileme ile Kanserdeki Moleküler Hedeflerin Hassas Tayini

Öz

Amaç: Belirli mutasyonlara neden olan moleküler değişiklikler tümör gelişimi ve sağkalımı için gereklidir. Bu moleküler hedeflerin doğru analiz edilmesi farklı kanser tiplerinde tanı, erken teşhis, prognoz tahmini ve tedavide yol gösterme açısından önemlidir. Bu nedenle, moleküler belirteçlerin hassas tayini için Sanger dizilemenin yanı sıra mini-dizileme protokolünü de optimize ederek kullanmayı amaçladık. Yöntem: Gliom örneklerinden elde edilmiş genomik DNAlar kullanılarak IDH1 R132, IDH2 R140/R172 ve TERT promotör C228/C250 mutasyonları için Sanger dizileme ve mini-dizileme yapılmıştır. Mini-dizileme reaksiyonları mutasyon saptama primerleri ile “SnaPshot Multiplex Ready Reaction Mix” kullanılarak gerçekleştirilmiş ve otomatik kapiler elektroforezinde yürütülmüştür. Çoklu eğriler “GeneMapper Software” kullanılarak analiz edilmiştir. Bulgular: Mini-dizileme analizlerinde, yabanıl tip alellere ve farklı mutasyonlara ait eğriler tespit edilmiştir. Yabanıl tip piklerin yanında yer alan eğriler, o noktada varyasyona işaret etmekte ve mutasyon renge göre tanımlanmaktadır. Sonuç: Moleküler belirteçlerin kanserde tayini oldukça önemlidir. Minidizileme yöntemi moleküler hedeflerin belirlenmesinde güvenilir bir yöntemdir.

Anahtar Kelimeler

Sensitive Detection of Molecular Targets in Cancer by Minisequencing

Abstract

Purpose: Molecular alterations leading to specific mutations are essential for tumor development and survival. Accurate analysis of these molecular targets is important for diagnosis, early detection, forecasting of prognosis and aiding in treatment of different cancer types. Therefore, for sensitive analysis of molecular markers, we aimed to optimize and use minisequencing protocols besides Sanger sequencing. Methods and Materials: Sanger sequencing and minisequencing were performed for IDH1 R132, IDH2 R140/R172 and TERT promoter C228/C250 mutations using genomic DNA isolated from glioma samples. Minisequencing reactions were performed with detection primers using SnaPshot Multiplex Ready Reaction Mix and run on automated capillary electrophoresis. Multiplex peaks were analyzed with GeneMapper Software. Results: In the multiplex minisequencing analyses, peaks corresponding to wild type alleles and different mutations were detected. The presence of the peaks next to the wild type peaks point to the presence of variations in that location and the nature of the mutation can be identified according to the color. Conclusions: Identification of molecular markers in cancer is very important. Minisequencing is a reliable method for the detection of molecular targets.

Keywords

References

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Details

Primary Language

English

Subjects

Biochemistry and Cell Biology (Other)

Journal Section

Research Article

Authors

Şirin Yüksel Kılıçturgay
Türkiye

Cemaliye Boylu Akyerli ^*
Türkiye

Publication Date

July 1, 2021

Submission Date

September 7, 2020

Acceptance Date

-

Published in Issue

Year 2021 Volume: 12 Number: 3

DOI

https://doi.org/10.31067/acusaglik.851006

IZ

https://izlik.org/JA23UM29YZ

Cite

RIS / Bibtex

EndNote

Yüksel Kılıçturgay Ş, Boylu Akyerli C (July 1, 2021) Sensitive Detection of Molecular Targets in Cancer by Minisequencing. Acıbadem Üniversitesi Sağlık Bilimleri Dergisi 12 3 547–552.