HiPure Cell miRNA Kit

Introduction The Kit is designed for purification of total RNA, including miRNA and other small RNA molecules (18nt), from cultured cells and various animal and human tissues, including difficult-to-lyse tissues samples. Details Specifications Features Specifications Main Functions Isolation total RNA and miRNA from cell and tissue without MagZol reagent Applications RT-PCR, Northern Blot, poly A purification, nucleic acid protection and in vitro translation Purification method Mini spin column Purification technology Silica technology Process method Manual (centrifugation or vacuum) Sample type Cell, Animal tissue, Plant tissue Sample amount Cells: ≤ 5 x 10^6, Animal tissue:<10mg Elution volume ≥30μl Time per run ≤40 minutes Liquid carrying volume per column 800µl Binding yield of column 100µg Principle Biological samples are first lysed and homogenized in a highly denaturing guanidine isothiocyanate-containing buffer, which immediately inactivates DNases and RNases to ensure isolation of intact DNA and RNA. The lysate is then passed through a Mini spin column. This column, in combination with the high-salt buffer, allows selective and efficient binding of genomic DNA. Flow-through from the column is digested by Proteinase K in the presence of ethanol. This optimized digestion, together with the subsequent addition of further ethanol, allows appropriate binding of total RNA, including miRNA, to the column. Contaminants are efficiently washed away and high-quality RNA is eluted. Advantages High quality – one step RNA extraction reagent combined with silica gel column can obtain the highest concentration Fast – several samples can be extracted in 40 minutes High applicability – samples including animals, plants, bacteria, cells, etc. High concentration – efficiently remove macromolecular RNA, enrich small RNA and improve sensitivity Safe – no phenol chloroform extraction and no use of Trizol reagent Kit Contents Contents R431102 R431103 Purification Times 50 Preps 250 Preps HiPure RNA Mini Columns 100 2 x 250 2ml Collection Tubes 100 2 x 250 Proteinase K 48 mg 240 mg Protease Dissolve Buffer 5 ml 15 ml Buffer RLC 40 ml 200 ml Buffer RWC 20 ml 80 ml Buffer RW2* 20 ml 2 x 50 ml RNase Free Water 10 ml 60 ml Storage and Stability Proteinase K should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions.