Comparative analysis of commercial qPCR master mixes for reliable plant telomere length measurement by monochrome multiplex qPCR

Şen Z. B., TURGUT KARA N.

Journal of Plant Biochemistry and Biotechnology, cilt.33, sa.1, ss.92-96, 2024 (SCI-Expanded) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 33 Sayı: 1
  • Basım Tarihi: 2024
  • Doi Numarası: 10.1007/s13562-023-00854-0
  • Dergi Adı: Journal of Plant Biochemistry and Biotechnology
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Agricultural & Environmental Science Database, BIOSIS, Biotechnology Research Abstracts, CAB Abstracts, Food Science & Technology Abstracts, Veterinary Science Database
  • Sayfa Sayıları: ss.92-96
  • Anahtar Kelimeler: Arabidopsis, Master mix, MMQPCR, Plant telomere, Telomere length measurement
  • İstanbul Üniversitesi Adresli: Evet

Özet

In the last decade, Monochrome Multiplex qPCR has been widely used for telomere length measurement studies. This study aimed to compare commercially available qPCR master mixes for relative plant telomere length measurement by Monochrome Multiplex qPCR and to facilitate optimization so that the technique can be used more widely and rapidly in plant studies. Relative telomere length was measured five different commercial qPCR master mixes. TelA and TelB primers were used to amplify Arabidopsis thaliana telomere sequences; single copy gene CYP5 was used as reference gene. Standard curves were generated for each master mix. T/S values were calculated by the ratio of Telomere quantity (T) to Single-copy gene quantity (S). It is found that different master mixes cause differences in Ct values and hence T/S ratios. Our results emphasized that certain master mixes may not be suitable for the MMQPCR technique or could yield varying outcomes.