A Technique for Magnetic Glass Particle-Mediated Nucleic Acid Extraction from Blood

Published: November 30, 2023

Abstract

Source: Rosenstierne, M. W., et al. Rapid, Safe, and Simple Manual Bedside Nucleic Acid Extraction for the Detection of Virus in Whole Blood Samples. J. Vis. Exp. (2018).

This video demonstrates a technique for nucleic acid extraction from whole human blood using magnetic glass particles (MGPs) — small magnetic beads coated with silica. Upon lysing the blood cells using a lysis buffer, the released nucleic acids bind to the surface of silica-coated MGPs, extracting the molecules from the sample.

Protocol

All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines 1. Blood Collection from Patients CAUTION: Take appropriate laboratory safety measures when collecting whole blood from the patient. Wear gloves and glasses. If the patient is in isolation, please follow biosafety level 4 procedures. To collect intravenous whole blood from the patient, use a butterfly need…

Açıklamalar

The authors have nothing to disclose.

Materials

Vacutainer K2 EDTA tubes (4 mL) Becton Dickinson 368861
Plus Blood Collection Becton Dickinson 362725
23G x 1 needle Becton Dickinson 300800
LUER LOK syringe (3 mL) Becton Dickinson 309658
Butterfly needle with small-bore extension tubing Jørgen Kruuse A/S 121714
1% Virkon Nomeco A/S 849265
MagNA Pure LC DNA Isolation Kit I – Lysis/Binding Buffer – Refill Roche Diagnostics A/S 3246752001
MagNA Pure LC Total Nucleic Acid Isolation Kit Roche Diagnostics A/S 3038505001
Nunc™ Biobanking and Cell Culture Cryogenic Tubes (1,8 mL) Thermo Fisher Scientific 375418
Nunc™ Biobanking and Cell Culture Cryogenic Tubes (3,6 mL) Thermo Fisher Scientific 379189
Nunc™ Biobanking and Cell Culture Cryogenic Tubes (4,5 mL) Thermo Fisher Scientific 379146
DynaMag™-5 Magnet Thermo Fisher Scientific 12303D
Transfer pipette (3.5 mL) Sarstedt 86.1171.010
Thermo Scientific™ Samco™ Fine Tip Transfer Pipettes (1.5 mL) Thermo Fisher Scientific 231
Thermo Scientific™ Nunc™ 50 mL Conical Sterile Polypropylene Centrifuge Tubes Thermo Fisher Scientific 339652

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Bu Makaleden Alıntı Yapın
A Technique for Magnetic Glass Particle-Mediated Nucleic Acid Extraction from Blood. J. Vis. Exp. (Pending Publication), e21842, doi: (2023).

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