The principle of the RINA- M14™ Viral Robotic Nucleic Acid Isolation Kit is optimized to RINA- M14™ robotic nucleic acid extraction system. First, in the presence of Proteinase K enzyme and with the presence of a concentrated chaotropic salt with CRNA and / or IC which were added by user and detergent mixture (Lysis Solution), any viral particles with lipid membrane or protein sheath (or viral or cell cultures containing IC ) is broken down and VNA occurs. Also in the presence of chaotropic salt and alcohol (BB Solution) added onto silica coated iron oxide nanoparticles (Magnetic Beads = MB) both IC and VNA are connected. Subsequently, with wash solution series in which the chaotropic salt reduced and alcohol is increased (WB1, WB2, and WB3) ICCs associated with MBs and VNAs are cleaned and dried. Robotic pipette tips are cleaned with Wash Solution after this step. Then, by washing with EB containing salt at low concentration, ICs and VNAs are passed through MB to the EB and the purification is completed. The CRNA acts as a crowding agent and increases the total amount of NA in the environment together with possible low-level target VNAs, which is necessary for NA purification. Because the purification of NA’s are very inefficient under a certain concentration. It also indirectly protects VNAs in the potential nuclease presence, some as targets for nuclease. IC is used to check if the VNA purification yield and subsequent Real Time PCR reaction inhibition are within certain limits. For example, PhCine Distemper Virus (PDV) when used in combination with the appropriate Bio-Speedy® Respiratory System Viral Factor Real Time PCR Panel such as “Bio-Speedy® Influenzae, Influenza B, Human RNazP Targeted Multiple 2X RT- qPCR Mixture” the result should be positive. A negative result may indicate a low in purification yield or an inhibition in the Real Time PCR reaction. It should not be forgotten that if DNA sequencing is done with the final VNA (Sanger, New Generation etc.), the results will be affected by the presence of CRNA and IC in the eluate.
INTENDED USE
RINA- M14™ Viral Robotic Nucleic Acid Isolation Kit is used for viral nucleic acid (VNA) isolation from nasal swabs, nasopharyngeal swabs, nasopharyngeal aspirate, bronchoalveolar lavage, sputum and other respiratory system samples (such as mucus-containing specimens to prevent pipetting, physical beating with glass beads, or incubation with dithiothreitol / n-acetyl-cysteine treatment is needed), mammalian sera, mammalian cell cultures and their media by using RINA ™ M14.