淋球菌中提取基因组DNA检测方案(核酸提取仪)

智能文字提取功能测试中

QuickGene DNA tissue kit S (DT-S)QuickGene SP kit DNA tissue (SP-DT)DF-5 Genomic DNA Extraction from Gonococcal Bacteria(Neisseria gonorrhoeae) Protocol Suspension of bacteria harvested from liquid medium after culture or agar medium 5，000 xg， 5 min → Remove supernatant Pelleted bacteria (about 5 mg of wet bacterial cell) MDT： 180pl · EDT： 20pl Suspend bacteria well by pipetting so as not to leave bacteria agglomerates Incubate at 55℃： 15 min*1 *1 If bacteria agglomerates remain at this time， break them —LDT： 180 ul by pipetting and incubate again. Vortex (maximum speed) ： 15 sec Flash spin down Incubate at 70℃：10 min Flash spin down >99% ethanol ： 240 pl Vortex (maximum speed)： 15 sec Flash spin down Transfer all contents of the micro tube into the cartridge of QuickGene Refer to the extraction protocol of the device used. QuickGene-810 to p.4-3 QuickGene-Mini80 to p.4-10 QuickGene SP kit to p.4-17 Genomic DNA (Elution volume： 200 pl) Results Bacterial strain ： Clinical isolates No.1~ 5 extracted from about 4.5 ~6 mg of each wet fungi Electropherogram Electrophoresis condition ： 1.5% agarose / 1 x TAE M： 入-Hind Ⅲ 1： Bacterial strain No.1 2： Bacterial strain No.2 3 ： Bacterial strain No.3 4 ： Bacterial strain No.4 5 ：Bacterial strain No.5 No decomposition was detected for extracted genomic DNA. |The yield of genomic DNA sample No.1 No.2 No.3 No.4 No.5 QuickGene 8.5 pg 7.1 pg 11.2 pg 11.0 pg 7.3 pg Spin column method (A company) 3.2 pg 6.6 pg 5.8 pg 6.5 pg 4.6 pg Protein contamination： A260/280 sample No.1 No.2 No.3 No.4 No.5 QuickGene 1.97 2.06 2.39 2.03 2.04 Spin column method (A company) 2.11 2.05 2.46 2.00 2.05 Chaotropic salt contamination ： A260/230 No Data Other · PCR ParC gene in subunit of topoisomeraselV as target of fluoroquinolone antibacterial agent was detected by PCR forgenomic DNA extracted using QuickGene system and Spin column method (A company). Electrophoresis condition ： 2% agarose / 1 x TAE M ： 100 bp DNA Ladder 1： Bacterial strain No.1 2 ： Bacterial strain No.2 3： Bacterial strain No.3 4 ： Bacterial strain No.4 5 ： Bacterial strain No.5 PCR products were detected for each genomic DNA. Common protocol is usable for the following Helicobacter pylori， Pseudomonas aeruginosa KKURABOThis book includes some protocol that have been performed， but not yet been approved. Depending on sample and storage conditions， nucleic acid maynot be extractable. Therefore， we cannot guarantee accurate data.The extracted nucleic acid contains unintended acid (ex： when extracting DNA， RNA is also extracted).-VII-· 淋球菌是一种严格的人体寄生菌，是淋病的病原体。提取淋球菌基因组DNA是一项便于开展后续分子生物学层面研究的基本工作。本文介绍采用Kurabo QuickGene核酸纯化系统及配套试剂对淋球菌中基因组DNA进行提取的实验方法。