The Extract Me RNA & DNA KIT is designed for a rapid, simultaneous isolation of high quality total RNA and genomic DNA from a single biological sample. Allows nucleic acids extraction from up to 30 mg of fresh or frozen tissue, and up to 107 cultured cells. It is ideal for studying transcriptome (RNA) and genome (DNA) in a single sample. The isolation protocols and buffer formulations were optimized for high isolation efficiency and purity of RNA and DNA. The product is intended for research use only.
The EXTRACTME RNA & DNA KIT utilizes spin minicolumns with membranes which efficiently and selectively bind nucleic acids at high concentration of chaotropic salts. During the first isolation step, a tissue is homogenized in order to disintegrate intercellular bonds (epithelial tissue) and fragmentize high-molecular proteins (muscle or connective tissue). A homogenate is lysed with guanidine thiocyanate and detergents. Nucleases are inactivated by guanidine thiocyanate and β-mercaptoethanol (optional). The homogenate is separated from undigested tissue/cell that remains after centrifugation. First, DNA selectively bounds to DNA Purification Column. In the second order, RNA remaining in the filtrate bounds to an RNA Purification Column membrane in the presence of additional ethanol. A three-step washing stage effectively removes impurities and enzyme inhibitors. Purified RNA and DNA are eluted with the use of low ionic strength buffers and may be used directly in all downstream applications, such as PCR, qPCR, primer extension, sequencing, microarrays, Southern blotting, and in case of RNA also Northern blotting, RT-PCR, cDNA synthesis or stored until ready to use.
- fresh or frozen tissue: up to 30 mg
- tissue preserved in RNase inactivating buffers (e.g. RNAlater® , Ambion):
up to 30 mg
- cell culture: up to 107 cells
~230 μg RNA, ~60 μg DNA
- 10 minutes (for simultaneous RNA/DNA purification, lysis and homogenization time not included)
- 15–30 minutes for homogenization in liquid nitrogen
- 15–20 minutes for mechanical homogenization (ceramic beads)
A260/A280 ratio = 1.9 – 2.1