Introduction This kit provides fast purification of high-quality RNA from cells, tissues, and yeast using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol/chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or isopropanol precipitation. RNA purified using the HiPure Total RNA Purification System can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation. Principle The Kit isolates total RNA from up to 107 cells or 20 mg tissue. A short workflow enables RNA isolation with genomic DNA removal in less than 25 mins. Samples are first lysed and homogenized. The lysate is passed through a DNA Mini column, ethanol is added to the flow-through, and the sample is applied to a RNA column. RNA binds to the membrane and contaminants are washed away. High-quality RNA is eluted in as little as 30µl water using the Kit. Advantages Efficient removal of DNA - unique genomic DNA removal column without DNase treatment High quality - high purity total RNA can be directly used in various sensitive downstream applications Fast - several samples can be extracted in 25 minutes by column method Safe - no phenol chloroform extraction required Sensitive - RNA can be recovered at the level of PG Details See protocol details in manual: Specifications Features Specifications Main Functions Isolation total RNA from 20mg tissue, 150mg plant, 5 x 106 cell using two columns (gDNA removed column) Applications RT-PCR, qRT-PCR, Northern hybridization, second generation sequencing Purification method Mini spin column Purification technology Silica technology, DNA filtration technology Process method Manual (centrifugation or vacuum) Sample type Animal soft tissue, cultured cells, lymphocytes, simple plant tissue Sample amount Cells:≤1 x 107 Animal tissue sample: 1-20 mg Plant tissue: 50-150 mg Yield 2-100μg Elution volume Variable to 50μl Time per run ≤25 minutes（1-24 samples） Liquid carrying volume per column 800µl Binding yield of column 100µg Kit Contents Contents R411102 D411103 Purification Times 50 Preps 250 Preps HiPure DNA Mini Columns 50 250 HiPure RNA Mini Columns 50 250 2ml Collection Tubes 100 2 x 250 Buffer RLC 40 ml 200 ml Buffer RW1 40 ml 200 ml Buffer RW2* 20 ml 2 x 50 ml RNase Free Water 10 ml 30 ml Storage and Stability HiPureKit can be stored dry at room temperature (15-25°C) and are stable for at least18 months under these conditions. During shipment, crystals or precipitation may form in the Buffer RLC. Dissolve by warming buffer to 37°C.
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