Q 100 droplet generator

Manufactured by Bio-Rad

Sourced in United Kingdom

The Q × 100 droplet generator is a laboratory instrument designed to generate monodispersed droplets for various applications. It provides a consistent and controlled method for producing uniform droplets with high precision.

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Lab products found in correlation

Q 100 droplet reader, by Bio-Rad (1 mentions) Ddpcr evagreen supermix, by Bio-Rad (1 mentions)

2 protocols using q 100 droplet generator

Allele-Specific ddPCR Quantification of rs1981760

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For allele-specific digital droplet PCR quantification of rs1981760-C compared with rs1981760-T we designed allele-specific probes (Supplementary Data 13), and performed amplification using the digital droplet PCR Supermix for Probes (BioRad) according to the supplied protocol. The probe for the T and C allele were designed to fluoresce in different channels allowing simultaneous detection of both in a single reaction. Droplets were generated on a q × 100 droplet generator (BioRad, UK) and droplets read on a q × 100 droplet reader. Probes targeting SOCS3 and JAK3 were used as positive controls for ChIP of STAT3, and RAB4 as a negative control. Input DNA as well as DNA obtained after ChIP was amplified on the same plate. For allele-specific wells, we performed technical duplicates.
The proportion of positive droplets relative to negative droplets is associated with the absolute concentration of the product according to a Poisson distribution. We therefore exploited this to empirically calculate the expected proportion of C versus T droplets (so as to take into account possible probe efficiency differences) and compared this to the actual ratio after ChIP. P-values using a χ²-test with two degrees of freedom were used to estimate the probability of the observed versus the expected ratio being due to chance.

Naranbhai V., Fairfax B.P., Makino S., Humburg P., Wong D., Ng E., Hill A.V, & Knight J.C. (2015). Genomic modulators of gene expression in human neutrophils. Nature Communications, 6, 7545.

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Droplet digital PCR for mtDNA quantification

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Each ddPCR reaction (V_F = 20 µL) contained 1× EvaGreen ddPCR Supermix (Bio-Rad), 100 nM of specific primers (Supplementary table S1) and DNA diluted to the equivalent of 1 oocyte/embryo. Prepared reactions were partitioned using a Q×100droplet generator (Bio-Rad) and the emulsions were transferred to a 96-well plate. PCR was performed at 95°C for 5 min, 50 cycles of 95°C, 57°C, and 72°C for 30 sec each, then held at 12°C. Fluorescence was measured using a Q×100droplet reader (Bio-Rad). The threshold for positive droplets was determined by the software and the final output was a concentration of starting template molecules/µL. Finally, the number of mtDNA copies or transcripts per reaction was normalized to obtain copies/per cell equivalent.

de Lima C.B., Martin H., Pecora Milazzotto M, & Sirard M.A. (2023). Genome-wide methylation profile of mitochondrial DNA across bovine preimplantation development. Epigenetics, 18(1), 2241010.

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