0000000000853893
AUTHOR
Ole Franz
DNA recovery from Droplet Digital™ PCR emulsions using liquid nitrogen
Droplet microfluidics is a technology that enables the production and manipulation of small volumes. In biosciences, the most popular application of this technology is Droplet Digital™ PCR (ddPCR™), where parallel nanoliter-scale PCR assays are used to provide a high sensitivity and specificity for DNA detection. However, the recovery of PCR products for downstream applications such as sequencing can be challenging due to the droplets' stability. Here we compared five methods for disrupting the droplets to recover DNA. We found that rapid freezing in liquid nitrogen results in a clear phase separation and recovery of up to 70% of the DNA content. Liquid nitrogen freezing can thus offer a si…
Retraction Note: Evaluating polymicrobial immune responses in patients suffering from tick-borne diseases
There is insufficient evidence to support screening of various tick-borne diseases (TBD) related microbes alongside Borrelia in patients suffering from TBD. To evaluate the involvement of multiple microbial immune responses in patients experiencing TBD we utilized enzyme-linked immunosorbent assay. Four hundred and thirty-two human serum samples organized into seven categories followed Centers for Disease Control and Prevention two-tier Lyme disease (LD) diagnosis guidelines and Infectious Disease Society of America guidelines for post-treatment Lyme disease syndrome. All patient categories were tested for their immunoglobulin M (IgM) and G (IgG) responses against 20 microbes associated wit…
Supplementary figure: DNA recovery from Droplet Digital PCR emulsions using Liquid Nitrogen
Supplementary figure 1 – Applicability of LN2 method to break ddPCR emulsion. ddPCR reactions were prepared with ddPCR™ Supermix for Probes (no dUTP) and Droplet Generation Oil for Probes (three tubes on the left) or ddPCR™ Supermix for EvaGreen® and Droplet Generation Oil for EvaGreen® (three tubes on the right). The tubes (A) and (D) show the emulsion before using LN2 method, and tubes (B), (C), (E) and (F) shows the oil and water layers after breaking the emulsion.
Supplementary table: DNA recovery from Droplet Digital PCR emulsions using Liquid Nitrogen
Supplementary table - Raw data from experiments studying DNA recovery from droplets using chemical and physical procedures.