Insight into DNA extraction of Candida sp : Comparison of an automated and A manual system

Sanjav Bhattacharya*. Pooja Pandey, H Anusha and Mammen Chandy

Author address: 

Tata Medical Center, South Lab, 14 Major Arterial Road (E-W), New Town, Rajarhat, Kolkata 700 156, India

Abstract: 

The objective of the study was to evaluate the efficacy of DNA extraction using automated and manual system. Both the methods were performed using Qiagen DNA Mini extraction kit. For automated DNA extraction, Qiacube (Qiagen) was used. Enzymatic lysis using lyticase (Sigma Aldrich) was carried out. Comparisons were made in terms of yield obtained and purity of the yield (using Thermo Scientific Nanodrop). Efficiency of extraction of the obtained product was checked in real time PCR (Rotorgene Q). Cycle threshold value obtained for the products in real-time PCR were compared. The Candida sp., used for the test were C. albicans, C. parapsilosis, C. tropicalis, C. famata, and C. dubliniensis. The results for the yield and purity obtained from both the methods were comparable. In terms of purity the 260/280 ratio were comparable whereas the 260/230 ratio showed some variation. Cycle threshold value in real-time PCR also showed similar values. Predesigned primers and probes (designed in lab) were used in the Real time PCR assay. Conclusion: No significant difference in manual and automated extraction using Qiacube for a member of Candida sp. was seen. The methods gave similar results in DNA yield and purity. Also, real-time amplification gave comparable values despite a difference in 260/230 ratio. 9830;Corresponding author E-mail: [email protected]
2014

abstract No: 

OP-011

Full conference title: 

Society for Indian Human and Animal Mycologists 2014
    • SIHAM