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

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.

1. To collect intravenous whole blood from the patient, use a butterfly needle with small-bore extension tubing and a blood collection vacuum tube containing a lysis/binding buffer. Rest the patient's arm in a downward position an....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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