Dergiler / Hacettepe Journal of Biology and Chemistry / 2020 / Cilt: 48 - Sayı: 1

COMPARISON OF BACTERIAL DNA EXTRACTION METHODS FROM STOOL SAMPLES FOR QUANTITATIVE REAL-TIME PCR ANALYSIS

Sayfa
41–48
Yayın Tarihi
2020-04-17
DOI
—
🏛️Bu makalenin bibliyografik verileri ve tam metin bağlantısı DergiPark üzerinden sağlanmıştır.
DergiPark Kaydı ↗
✨

Yapay Zeka Hızlı Bakış & Çıkarımlar

AI Insight
⚡ TL;DR(1 Cümlede Hızlı Çıkarım)

Commercial kits have better results in terms of DNA recovery and purity, and have more accurate results for qPCR experiments.

📌 3 Maddede Bu Makale

🎯
Amaç & Hipotez:The most crucial step in molecular microbiology studies is choosing an appropriate procedure to obtain sufficient, high-quality DNA.
📊
Temel Bulgular & Veri:DNA yield was significantly higher in Invitrogen when compared to phenol-chloroform method.
💡
Sonuç & Katkı:Commercial kits have better results in terms of DNA recovery and purity, and have more accurate results for qPCR experiments.
🔬Tür:Araştırma Makalesi
📍Kapsam:Genişletilmiş Akademik Örneklem
⚙️Değişkenler:qPCR · DNA extraction · stool

Abstract

The most crucial step in molecular microbiology studies is choosing an appropriate procedure to obtain sufficient, high-quality DNA. Two commercially available kits (QIAamp DNA Stool Mini Kit and PSP Spin Stool DNA Plus Kit) and a modified in-house phenol-chloroform extraction method was compared for their efficiency in isolation of certain bacteria from human stool samples for Real-Time qPCR. DNA yield was significantly higher in Invitrogen when compared to phenol-chloroform method. Spiking experiments and qPCR results revealed that efficiency was in phenol-chloroform extraction was lower when compared to other two commercial kits. Commercial kits have better results in terms of DNA recovery and purity, and have more accurate results for qPCR experiments.

🔖

Bu Makaleye Atıf Yap

KOCAK, C. O. (2020). COMPARISON OF BACTERIAL DNA EXTRACTION METHODS FROM STOOL SAMPLES FOR QUANTITATIVE REAL-TIME PCR ANALYSIS. Hacettepe Journal of Biology and Chemistry, Cilt: 48 - Sayı: 1, 41–48. https://ajindex.com/articles/255915

📄 Tam Metin PDF Okuyucu

📄

PDF görüntüleyici hazırlanıyor...

Kaynakça (1)

  1. Turnbaugh PJ, Ley RE, Hamady M, Fraser-Liggett CM, Knight R, Gordon JI, The Human Microbiome Project, Nature, 449 (2007) 804-810.2. Guarner F, Malagelada JR, Gut flora in health and disease, Lancet, 361 (2003) 512-519.3. Abt MC, Artis D, The intestinal microbiota in health and disease: the influence of microbial products on immune cell homeostasis, Curr Opin Gastroenterol, 25 (2009) 496-502.4. Bokulich NA, Chung J, Battaglia T, Henderson N, Jay M, Li H, A DL, et al., Antibiotics, birth mode, and diet shape microbiome maturation during early life, Sci Transl Med, 8 (2016) 343ra382.5. Turnbaugh PJ, Hamady M, Yatsunenko T, Cantarel BL, Duncan A, Ley RE, Sogin ML, et al., A core gut microbiome in obese and lean twins, Nature, 457 (2009) 480-484.6. Turnbaugh PJ, Ley RE, Mahowald MA, Magrini V, Mardis ER, Gordon JI, An obesity-associated gut microbiome with increased capacity for energy harvest, Nature, 444 (2006) 1027-1031.7. Weinstock GM, Genomic approaches to studying the human microbiota, Nature, 489 (2012) 250-256.8. Mardis ER, Next-generation DNA sequencing methods, Annu Rev Genomics Hum Genet, 9 (2008) 387-402.9. Ley RE, Backhed F, Turnbaugh P, Lozupone CA, Knight RD, Gordon JI, Obesity alters gut microbial ecology, Proceedings of the National Academy of Sciences of the United States of America, 102 (2005) 11070-11075.10. Sekirov I, Russell SL, Antunes LC, Finlay BB, Gut microbiota in health and disease, Physiol Rev, 90 (2010) 859-904.11. Poretsky R, Rodriguez RL, Luo C, Tsementzi D, Konstantinidis KT, Strengths and limitations of 16S rRNA gene amplicon sequencing in revealing temporal microbial community dynamics, PLoS One, 9 (2014) e93827.12. Rintala A, Pietila S, Munukka E, Eerola E, Pursiheimo JP, Laiho A, Pekkala S, et al., Gut Microbiota Analysis Results Are Highly Dependent on the 16S rRNA Gene Target Region, Whereas the Impact of DNA Extraction Is Minor, J Biomol Tech, 28 (2017) 19-30.13. Guo X, Xia X, Tang R, Zhou J, Zhao H, Wang K, Development of a real-time PCR method for Firmicutes and Bacteroidetes in faeces and its application to quantify intestinal population of obese and lean pigs, Lett Appl Microbiol, 47 (2008) 367-373.14. Ott SJ, Musfeldt M, Ullmann U, Hampe J, Schreiber S, Quantification of intestinal bacterial populations by real-time PCR with a universal primer set and minor groove binder probes: a global approach to the enteric flora, J Clin Microbiol, 42 (2004) 2566-2572.15. Fouhy F, Clooney AG, Stanton C, Claesson MJ, Cotter PD,