试剂盒提取详细步骤.docVIP

美国MOBIO土壤RNA提取试剂盒(RNA PowerSoil? Total RNA Isolation Kit) Detailed Protocol (Describes what is happening at each step) Wear RNase-Free gloves (1555) at all times and remove RNase from the work area using Lab Cleaner (12095) for RNase Removal. Both of these products are available from MO BIO. Please see “Other Quality Products Available” section at the end of this manual. 1. Add up to 2 g of soil to the 15 ml Bead Tube (provided). Note: Please refer to Hints and Troubleshooting Guide for information regarding the amount of soil to process. 1、加2g土壤到15ml磁珠管中。 注意：请参照提示和纠错指南确定加入土壤的量。 2. Add 2.5 ml of Bead Solution to the Bead Tube and vortex to mix. 2、加2.5ml Bead Solution到磁珠管中并漩涡混合。 3. Add 0.25 ml of Solution SR1 to the Bead Tube and vortex to mix. What’s happening: The Bead Solution is a buffer used to disperse cells and soil particles. Solution SR1 contains SDS and other disruption agents which aid in complete cell lysis. In addition to aiding in cell lysis, SDS is an anionic detergent that breaks down fatty acids and lipids associated with the cell membrane of several organisms. Note: If it gets cold, it will form a white precipitate. Heating to 60oC will dissolve the SDS and will not harm the other disruption agents. 3、加0.25ml SR1到磁珠管并漩涡混合。 发生的反应：Bead Solution是一种缓冲液可用来打散细胞和土壤颗粒。 SR1 包括SDS和其他的能帮助细胞裂解的破碎剂。除了帮助细胞破碎外，SDS是阴离子洗涤剂，可以破坏脂肪酸和几种微生物细胞膜相关的脂类。 注意：如果天气冷了，可能会形成白色沉淀。加热到60℃溶解SDS，且不会影响其他裂解试剂的作用。 4. Add 0.8 ml of Solution SR2 and place the Bead Tube on the Vortex Adapter (MO BIO Catalog # 13000-V1-15 for Vortex Genie 2 or 13000-LV2-15 for Labnet Vortex) and vortex at maximum speed for 5 minutes. What’s happening: Solution SR2 is a precipitation reagent used to remove non-DNA organic and inorganic material including humic substances, cell debris, and proteins. Contaminating organic and inorganic matter may reduce DNA purity and inhibit downstream DNA applications. Vortexing is critical for homogenization and cell lysis. 4、加0.8ml SR2到磁珠管，最大转速旋转5min。 反应：SR2是一种沉淀试剂，用来去除非DN