Omega Bio-Tek/Mag-Bind® cfDNA Kit/5-preps/M3298-00

Overview

The Mag-Bind® cfDNA Kit is designed for rapid and reliable isolation of circulating DNA from 500-8,000 µL plasma or serum samples. The Mag-Bind® cfDNA Kit can be processed manually or on an automated platform. The procedure eliminates the need for funnels and vacuum steps, providing hands-free operation in automated protocols.

The uniquely formulated binding buffer allows for large sample volumes to be processed in automated formats which allow for 4 mL of serum or plasma to be processed in a single well in 24-well plate format. The magnetic response of the Mag-Bind® Particles CH allows for fast magnetization during steps requiring high volumes, and the high binding capacity allows for reduced amounts of magnetic particles required, thus reducing the elution volume down to 50 µL for 4mL serum or plasma samples.

The Mag-Bind ® cfDNA Kit has also been automated to process 8 mL samples on Hamilton platforms. Please contact us for automation support.

Protocols are available for the following automated platforms:

• Hamilton Microlab® STAR
• Hamilton Microlab® NIMBUS
• KingFisher™, BioSprint®, and MagMAX® 96

Protocol and Resources

Resources & Literature

Product Data

Cell-free DNA purified using Omega Bio-tek’s Mag-Bind ® cfDNA Kit has little genomic DNA carryover

Figure 1. Electropherogram overlay of purified DNA from 4 mL serum. 4 mL of unspiked serum was purified using kits from Omega Bio-tek and Company Q following manufacturer’s recommended protocols. Purified DNA was analyzed on Agilent’s TapeStation® 2200.

High quality and yield for cell-free DNA purified using Omega Bio-tek’s Mag-Bind ® cfDNA Kit Mag-Bind® cfDNA kit

Figure 2.  Electropherogram overlay of purified DNA from 1 mL serum. Sheared, bacterial DNA was spiked into serum samples and isolated using Omega Bio-tek’s Mag-Bind cfDNA Kit and a column-based kit from Company Q. Real-time PCR was performed using 16s bacteria-specific primers at 1X and 10X dilutions. Purified DNA was analyzed on Agilent’s TapeStation® 2200. The Omega Bio-tek kit was able to capture the circulating, cell-free DNA with no genomic DNA contamination. In contrast, Company Q’s eluate contained high molecular weight fragments indicating the presence of genomic DNA in the circulating DNA isolation.

cfDNA purified using Omega Bio-tek’s cfDNA kit contains no PCR inhibitors

Table 1. Real-time PCR with 16S bacterial-specific primers was performed on triplicates of undiluted and 10-fold dilutions of DNA isolated from samples purified [figure 2]. The data shows the purified DNA is free of PCR inhibitors and the specific cfDNA concentration is higher than Company Q.

Circulating, cell-free DNA as small as 50 bp can be recovered with Mag-Bind® cfDNA Kit

Figure 3. DNA fragments as small as 50 bp can be recovered using modified protocol. Serum spiked with 50 bp DNA ladder was purified with the Mag-Bind® cfDNA Kit. By using modified protocols, smaller fragments were recovered.

Analysis of cfDNA purified from 8 mL plasma using Mag-Bind® cfDNA Kit

Figure 4.1. Plasma samples were purified using the Mag-Bind® cfDNA Kit. The purified DNA samples were loaded on a D100 high-sensitive tape on TapeStation® 2200.

Figure 4.2. Gel image from figure 4.1 was analyzed using the TapeStation software. Peak and concentration data showed that 8 mL automation process was a success.

Figure 4.3.  Gel image from figure 4.1 was analyzed using the TapeStation software. The overlay showed that the purified DNA contained more cfDNA and minimal genomic DNA contamination.

Citations