抽plasmiddna.pptVIP

A rapid method for making a small preparation of purified plasmid DNA is known as a Plasmid Miniprep Purification Kit . 工作流程 The Cell Resuspension Solution contains Tris (a buffering agent), EDTA, and RNase. EDTA stabilizes the cell membrane by binding the divalent cations of Mg++ and Ca++. RNase destroys the cell’s RNA. The resuspended cells are then treated with a mixture of sodium dodecyl sulfate (SDS) and sodium hydroxide. SDS, an ionic detergent, dissolves the phospholipid and protein components of the cell membrane. This lyses the membrane releasing the cell contents. Sodium hydroxide denatures both plasmid and chromosomal DNAs into single strands. The chromosomal DNA separates completely into individual strands; however, the single-strained plasmid loops remain linked together like interlocked rings. Treatment with potassium acetate and acetic acid forms an insoluble precipitate of SDS/lipid/protein and neutralizes the sodium hydroxide from the previous step. At neutral pH, the DNAs renature and become trapped in the SDS/lipid/protein precipitate. The plasmid DNA completely renatures into double-stranded molecules that remain in solution The precipitate is pelleted by centrifugation and discarded, leaving the plasmid DNA (as well as small RNA molecules) in the supernatant. The supernatant is collected and purified by binding to glass fines. Isolation and Analysis of Recombinant Plasmids Isolation and Analysis of Recombinant Plasmids Isolation and Analysis of Recombinant Plasmids Isolation and Analysis of Recombinant Plasmids Isolation and Analysis of Recombinant Plasmids * * 定序工作流程介紹 邱瓊瑩 收件、分類 養菌、抽plasmid DNA 純化PCR product 電泳分析 定序反應 酒精沉澱 上3730 分析報告 1.拆包裝(宅配) 2.清點數量(樣品與primer) 3.填上訂單編號(宅配的收據單上也要填寫) 4.樣品袋上填上訂單編號、客戶單位、聯絡人 5.登記在收件本上 6.影印2份(一份放置櫃檯，一份放置業務助理) 順序由訂單編號由大至小排列(號碼大的在上方) 7.將樣品分類放置 菌 菌的箱子, 菌的primer 菌的primer的箱子 DNA, PCR product 收件箱, 上機 客戶外送上機盒子 8.將訂單分類放好 菌 養菌單, 上機 直接上機 DNA, PCR product