Briefly; first of all the user is allowed to homogenize and lyse the tissues outside the robot, resulting in cleaned and liquefied lysate. Then nucleic acid purification of this lysate is carried out in the RINA-M14 ™ robot with RINA-14™ Blood and Tissue Robotic Nucleic Acid Isolation Kit. The details of the working principle are as follows: first, tissues (IC can be added if desired by the user) are homogenized with a vortex or homogenizer in the presence of detergent and PCR inhibitor eliminating agent solution (TL Solution). During homogenization, the tissues and microorganisms are completely or partially lysed depending on their type. Next, if a homogenizer is used, to get rid of the homogenization consumables in the tube such as the bead etc. the homogenate is centrifuged at 3,000 g for 1 second, and supernatant is transfered into a new tube. The physical lysis process is then continued by heat treatment. Subsequently the enzymatic (Proteinase K) lysis of the homogenate / lysate is completed. The tissue is now lysate. The lysate is removed by centrifugation. Both NA’s and IC (if IC added) are binding to silica coated iron oxide microparticles (Magnetic Bead Solution = MB Solution) in the presence of concentrated chaotropic salt, detergent mixture (LB Solution (IC can be added if requested by the user)), and alcohol (Solution B). Then the ICs and NAs bound to the MBs with the wash solution series (W1, W2 and W3) with reduced chaotropic salt and increased alcohol are cleaned, dried and rinsed with the MBR Solution. . After this step the robotic pipette tips are cleaned once again with TR Solution (at the stage of provocation). Subsequently, the solution is washed with E solution containing salt at low concentration, and the purification is completed by passing the ICs and NAs through the MB solution to the E solution. Finally, if DNA sequencing is to be performed with the obtained NA (Sanger, New Generation, etc.) it should be considered whether or not there is IC in the eluate.
INTENDED USE
The kit is used RINA ™ M14 fully automated for host genomic DNA, bacterial, parasite etc. microbial and viral nucleic acid purification from animal tissues including blood.