The MasterPure™ Complete DNA and RNA Purification Kit, MasterPure™ DNA Purification Kit, and MasterPure™ RNA Purification Kit use a novel, patent-pending technology that enables efficient purification of intact, high molecular weight DNA or RNA from a wide range of biological materials. Total nucleic acid (TNA), DNA, or RNA can be purified from bacteria, tissue culture cells, frozen or fresh mammalian tissue, paraffin-embedded tissue sections, mouse tail snips, and other biological materials. The protocol for TNA purification involves (1) an enzyme/salt/detergent-mediated cell lysis, (2) protein removal by precipitation, and (3) nucleic acid precipitation and resuspension. To isolate purified DNA, the sample is treated with RNase A; to isolate purified RNA, the sample is treated with RNase-Free DNase I. All manipulations are performed in standard microcentrifuge tubes. The recovered nucleic acids have an A260/280 ratio of 1.8-2.0, and can be used directly in PCR amplification (DNA and TNA) or RT-PCR (RNA). The standard MasterPure™ Complete DNA and RNA Purification Kit contains sufficient reagents for 200 DNA purifications or 100 DNA and 100 RNA purifications.
|Human & Mammalian
|RNA & Genomic DNA
||Tissue culture cell lines
Table 1. Examples of targets and samples analyzed using MasterPure™ DNA and RNA Purification Kits. Using an extremely simple salt precipitation protocol, TNA, DNA, or RNA can be isolated from virtually any source in less than 1 hour.
Figure 1. DNA, RNA, and total nucleic acid (TNA) purified from diverse cell sources using the MasterPure™ Complete Kit. M = kb ladder.
Figure 2. Basic MasterPure™ Complete Kit extraction protocol.
|Sample||Sample Size||TNA µg||DNAµg||RNA µg|
||1 X 106 cells
||3.5 x 106 cells
||2.2 x 106 cells
||1.1 x 107 cells
Table 2. Example of yields of nucleic acids obtained using MasterPure™ Complete DNA and RNA Purification Kits.
*For extraction of yeast DNA, the MasterPure Yeast DNA Purification Kit is recommended.
Figure 3. Consistent purification of DNA, total nucleic acid, and RNA. Five different genomic DNA (Lanes 1-5), total nucleic acid (Lanes 6-10), and total cellular RNA (Lanes 11-15) samples were isolated from frozen bovine liver with the MasterPure™ Complete DNA and RNA Purification Kit. M = kb ladder.
Figure 4. Comparison of PCR sensitivity using templates obtained using the MasterPure™ Complete DNA and RNA Purification Kit versus a spin column kit. PCR amplification was performed after extraction from the indicated number of E. coli cells.
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