Briefly; the user first obtains feces lysate liquefied and cleaned by homogenization, lysing and performing PCR inhibitor elimination outside of the robot Then nucleic acid purification of this lysate is carried out in the RINA-M14 ™ robot which uses RINA- M14™ Stool Robotic Nucleic Acid Isolation Kit. The details of the working principle are as follows: the feces (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 (FL Solution). During homogenization, the feces 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 feces is now lysate. The lysate is removed by centrifugation. Both NA’s and IC if 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
RINA- M14™ Stool Robotic Nucleic Acid Isolation Kit is used RINA ™ M14 fully automated for host genomic DNA, bacterial, parasite etc. microbial and viral nucleic acid purification from mammalian feces sample.