Reagent for removing non-nuclear red blood cells in biological sample and application of reagent to DNA extraction
A technology for biological samples and nucleated red blood cells, applied in DNA extraction, and in the field of lysing reagents for removing anucleated red blood cells from biological samples, can solve the problems of adverse effects of extraction or culture, incomplete lysis of red blood cells, and low purity of nucleated cells , to achieve the effects of safe and environmentally friendly reagent preparation and use, convenient downstream genetic manipulation detection, and chemical stability of reagent components
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Example Embodiment
[0031] Example 1: Preparation of erythrocyte lysis reagent
[0032] According to the content of the invention, four erythrocyte lysis reagents A to D were prepared, and the cell lysis reagent E (ACK ammonium chloride erythrocyte lysis solution) was prepared as a control, as follows:
[0033] (1) Erythrocyte Lysis Reagent A
[0034] EDTA 5mmol / L, NaCl 15 mmol / L, Tween 20 0.4% (v / v), Tris-HCl 5 mmol / L, pH about 8.0.
[0035] (2) Erythrocyte Lysis Reagent B
[0036] NTA 3mmol / L, NaCl 50 mmol / L, Triton X-100 0.8% (v / v), Tris-HCl 15 mmol / L, pH about 7.6.
[0037] (3) Erythrocyte Lysis Reagent C
[0038] EDTA 1mmol / L, KCl 40 mmol / L, Triton X-100 0.3% (v / v), Tris-HCl 20 mmol / L, pH about 7.5.
[0039] (4) Erythrocyte Lysis Reagent D
[0040] EDTA 2mmol / L, NaCl 35 mmol / L, Triton X-100 0.5% (v / v), Tris-HCl 10 mmol / L, pH of the reagent is about 7.8.
[0041] (5) Erythrocyte Lysis Reagent E
[0042] NH 4 Cl 155 mmol / L, KHCO 3 12mmol / L, EDTA 0.15mmol / L, pH7.2-7.4.
[0043] After p...
Example Embodiment
[0044] Example 2: Comparison of the lysis and removal effects of different formulations of cell lysis reagents on peripheral blood red blood cells
[0045] (1) Sample collection: Collect 1 aliquot of EDTA anticoagulated whole blood with a volume of 5 ml, and after mixing, dispense 400 μl / tube into 2 ml centrifuge tubes, and dispense 10 tubes of samples in total.
[0046] (2) Take the five erythrocyte lysis reagents prepared in Example 1, according to the ratio of whole blood: erythrocyte lysis solution volume to 1:3, draw 1.2 ml of each erythrocyte lysis reagent and add it to the divided whole blood, each erythrocyte lysis solution. Lyse 2 tubes of samples, a total of 5 groups of 10 samples, and mix thoroughly for 1 min.
[0047] (3) Centrifuge 5 groups of 10 tubes at 10,000 rpm for 1 min, discard the supernatant to retain the enriched cell pellet.
[0048] (4) Add 1 ml of erythrocyte lysis reagent to the enriched cell pellet, mix well and resuspend the pellet.
[0049] (5) ...
Example Embodiment
[0054] Example 3: DNA extraction application of cell lysis reagents in Epstein-Barr virus positive whole blood samples
[0055] (1) DNA extraction solution: prepared according to the formula of 10mmol / L NaOH and 0.1% SDS for DNA extraction.
[0056] (2) Sample collection: There are various related diseases caused by human infection with EB virus, which first proliferate in oropharyngeal epithelial cells, then infect B lymphocytes, enter the blood circulation to cause systemic infection, and can be latent in human lymphoid tissue and white blood cells for a long time. . The present invention collects 5 cases of EDTA anticoagulant human whole blood samples identified as Epstein-barr virus (Epstein-barr virus) positive from the clinic, and each case is divided into 400μl / tube and placed in a 1.5ml centrifuge tube, and divided into 2 groups with a total of 10 Tube.
[0057] (3) Take erythrocyte lysing reagent D and control erythrocyte lysing reagent E in Examples 1 and 2, which ...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap