DNA Extraction From Whole Blood on Hamilton STAR

High yield and quality from 1 mL whole blood in less than 2 hours automated

Download this article here.

Introduction

Nucleic acid extraction from whole blood is the initial step for many genomic-based diagnostic workflows including biomarker discovery, newborn screening and pharmacogenetics. Extracting large amounts of high quality genomic DNA is crucial for downstream applications such as qPCR, microarray analysis and next generation sequencing. This creates the need for reliable extraction methods from large volume blood samples (> 250 µL) which can be extremely demanding and time consuming, thus creating bottlenecks within processes.

Omega Bio-tek's Mag-Bind® Blood DNA HDQ 96 Kit offers a solution for such difficult procedures and has been successfully validated on Hamilton's Microlab STAR. The system uses a completely automated protocol, which improves accuracy by eliminating errors than can occur during manual processing, and can extract pure DNA from 96 1 mL whole blood samples in under 2 hours.

Hamilton® Microlab STAR Deck Layout


Materials and Methods

DNA was extracted from two sets of identical whole blood samples on the same day. One set was processed using Company Q's automated system with Company Q's chemistry, and one set was processed using the Hamilton Microlab STAR and Omega Bio-tek's Mag-Bind® Blood DNA HDQ 96 Kit. For the Hamilton/Omega procedure, whole blood tubes were inserted into sample carriers where the instrument scanned barcodes from the individual blood tubes. 1,000 µL of each blood sample was then aspirated and dispensed into 250 µL aliquots in the same well position of 4 different 96-well deep well plates. Cell lysis was achieved by heating and shaking the samples using the Hamilton heater shakers. Magnetic beads and binding solution were then added to capture the DNA from the sample lysates. Four quick wash steps were performed and then DNA was eluted in 10 mM Tris-Cl (pH 8.5). No drying step was required to remove ethanol allowing for faster processing and more reliable results. DNA purity was analyzed with the Thermo Scientific NanoDrop 2000c and quantity was assessed with Promega QuantiFluor dsDNA system.

Intrument Configuration Specifications
Hamilton equipment
  • Hamilton Microlab STAR configured with 96-channel head
  • 8-channel independent pipetting heads
  • Heater shaker units
  • iSWAP arm
  • Liquid waste module
Plasticware
  • Nalgene Nunc 96-well plates (278752)
  • Hamilton 1,000 µL tips (182085)
  • Hamilton 300 µL reservoir (56669-01)
Reagents

Genomic DNA extracted from 1 mL whole blood on the Hamilton STAR with Omega Bio-tek chemistry. DNA was analyzed with the Thermo Scientific NanoDrop 2000c.

Total processing time reported from customer for 96 1 mL whole blood samples from blood tubes to purified DNA.
Results

Automation of genomic DNA purification from whole blood on the Hamilton Microlab STAR with Omega Bio-tek's chemistry allows for recovery of large amounts of high quality genomic DNA. The Hamilton/Omega procedure decreased overall processing time by simultaneously extracting from 96 samples compared to batches of 24 processed by Company Q. The Hamilton/Omega instrument scripting has been optimized to allow for maximum throughput, DNA quality and yield. Minimal user intervention is required as the system goes from whole blood tubes to purified DNA without any user pause steps after initial inputs are received.

Sample Yield (µg) Yield (µg)
  Omega Bio-tek Company Q
1 49.69 18.0
2 30.88 31.1
3 18.26 10.0
4 25.16 20.8
5 38.55 29.1
6 33.58 29.1
7 33.58 29.1
8 25.39 21.3
9 37.34 26.4
10 53.87 25.8
Average 33.67 24.1

Genomic DNA extracted from 1 mL whole blood on the Hamilton STAR with Omega Bio-tek chemistry. DNA was analyzed with Promega QuantiFluor dsDNA Kit.
Ordering Information
Description Product No Size

Mag-Bind® Blood & Tissue DNA HDQ Kit

M6399-01 4 x 96