Lysis binding solution for rapidly extracting RNA by paramagnetic particle method, kit and application thereof

[0034]Example 1: Two different levels of the reagent formulation of the present invention

[0035]1. Preparation of more fruitful (SIGMA, Item No. 45466, powder), sodium chloride (Sigma, Item No. S3014, granules), sodium citrate (SIGMA, Item number C8532, granular), DTT (Solarbia, Item No. 578517, Particles The article), EDTA-2NA (Sigma, Item No. E7889, Liquid, 0.5m), Polyethylene glycol 8000 (PEG8000, Sigma, Item No .: P5413, Powder), Proclin300 (Solarbio, Liquid, 100%), and other configuration Need H2O, vessel or consumables, etc .;

[0036]2, according to a certain ratio, the above reagent is configured as a corresponding mother liquor:

[0037] Reagent Mother liquor concentration More fruitful 20mm Sodium chloride 5M Sodium citrate 0.1M DTT 1M EDTA-2NA 0.5m (original concentration) PEG8000 50% (w / v)

[0038]3, configure the lysate: Add the reagent separately according to the following formulation

[0039] Reagent name (mother liquor) Cracked liquid formula 1 Cracked liquid formula 2

[0045]Example 2: Reagent Properties of the present invention

[0046]1. Sample preparation: Prepare fake viruses to extract, and inactivate the fake virus 56 ° C, 30 min. After inactivation, dilute the fake virus to 200 copies / ul, ready for RNA extraction.

[0047]2, nucleic acid extraction: Add 200 ul of the inactivated pseudogenic toxic to the dispensed formulation 1 reactive plate and formulation 2 reactive plate reagent board, according to the "extraction procedure" described below, using the automated extractor, After the extraction, the eluate was collected and used for subsequent PCR reactions.

[0048]Extract

[0049] name time Module temperature Magnetic bead mix 20sRoom temperature Shield 40sRoom temperature Cracking combination 360s90℃ washing 40s80℃ washing 40s80℃ washing 40s80℃ Elution recovery 300s85℃ Abandon20sRoom temperature

[0050]3, the nucleic acid extracted by the recipe 1 reaction plate is labeled as sample 1; the nucleic acid extracted by the recipe 2 reaction plate

[0053]Example 3: Competitive performance verification

[0054]1. Sample preparation: Prepare fake viruses to extract, and inactivate the fake virus 56 ° C, 30 min. After inactivation, dilute the fake virus to 200 copies / ul, ready for RNA extraction.

[0055]2, nucleic acid extraction: Add 200UL after the inactivated pseudogenic toxic to the Sub-Socrat reactive plate, according to the "extraction procedure" described below, use the automated extractor to extract, after the extraction is completed, The eluate is collected for subsequent PCR reactions.

[0056]Extract

[0057] name time Module temperature Magnetic bead mix 20sRoom temperature Shield 40sRoom temperature Cracking combination 360s90℃ washing 40s80℃ washing 40s80℃ washing 40s80℃ Elution recovery 300s85℃ Abandon20sRoom temperature

[0058]3, the resulting nucleic acid labeled sample ZK.

[0059]4, PCR reaction: Take sample ZK5UL, add 15 ul of amplification reagent for PCR reaction, each reaction is repeated 3 times, according to the