Accurate determination of DNA yield from individual mosquitoes for population genomic applications  

作　　者：Craig S. Wilding D. Weetman K. Steen M.J. Donnelly 

机构地区：[1]Vector Group, Liverpool School of Tropical Medicine, Pembroke Place, Liverpool, L3 5QA, UK

出　　处：《Insect Science》2009年第4期361-363,共3页昆虫科学（英文版）

摘　　要：Accurate estimates of DNA quantity are likely to become increasingly important for successful genomic screening of insect populations via recently developed, highly multiplexed genotyping assays and high-throughput sequencing methods. Here we show that genomic DNA extractions from single Anopheles gambiae Giles using a standard commercial kit-based methodology yield extracts with concentrations below the linear range of spectrophotometric absorbance at 260 nm. Concentrations determined by spectropho- tometry were not reproducible, and are therefore neither accurate nor reliable. However, DNA quantification using a fluorescent nucleic acid stain （PicoGreen） gave highly reproducible concentration estimates, and indicated that, on average, single mosquitoes yielded approximately 300 ng of DNA. Such a total yield is currently insufficient for many highthroughput genome screening applications, necessitating whole genome amplification of all or most individuals in a population prior to genotyping.Accurate estimates of DNA quantity are likely to become increasingly important for successful genomic screening of insect populations via recently developed, highly multiplexed genotyping assays and high-throughput sequencing methods. Here we show that genomic DNA extractions from single Anopheles gambiae Giles using a standard commercial kit-based methodology yield extracts with concentrations below the linear range of spectrophotometric absorbance at 260 nm. Concentrations determined by spectropho- tometry were not reproducible, and are therefore neither accurate nor reliable. However, DNA quantification using a fluorescent nucleic acid stain （PicoGreen） gave highly reproducible concentration estimates, and indicated that, on average, single mosquitoes yielded approximately 300 ng of DNA. Such a total yield is currently insufficient for many highthroughput genome screening applications, necessitating whole genome amplification of all or most individuals in a population prior to genotyping.

关 键 词：Anopheles gambiae DNA quantification MOSQUITO PICOGREEN SPECTROPHOTOMETRY 

分 类 号：Q959.212.06[生物学—动物学] Q959.468