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Sybrselect for cfx master mix

Manufactured by Thermo Fisher Scientific
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SYBRSelect for CFX Master Mix is a ready-to-use qPCR reagent designed for use with the CFX platform. It contains SYBR Green I dye for detection of PCR amplification.

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Lab products found in correlation

Cfx384 thermocycler, by Thermo Fisher Scientific (2 mentions)

2 protocols using sybrselect for cfx master mix

1

Quantitative PCR of 16S rRNA genes

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Quantitative PCR (qPCR) was performed on 216/290 samples that were available from the 16S rRNA gene sequencing cohort. qPCR of total 16S rRNA gene copies was carried out in triplicate 10 μL reactions with 200 nM 340F/514R primers using a BioRad CFX384 thermocycler with SYBRSelect for CFX Master Mix (Life Technologies) according to the manufacturer’s instructions and an annealing temperature of 60 °C. Absolute quantifications were determined based against a standard curve of purified 8F/1542R amplified E. coli MG1655 DNA42 . Reactions were performed in triplicate and mean values were taken for further downstream analyses. Absolute bacterial abundance was derived by adjustments for dilutions during DNA extraction, normalization, and PCR reaction preparation dividing by the total fecal mass used for DNA extraction in grams.
von Schwartzenberg R.J., Bisanz J.E., Lyalina S., Spanogiannopoulos P., Ang Q.Y., Cai J., Dickmann S., Friedrich M., Liu S.Y., Collins S.L., Ingebrigtsen D., Miller S., Turnbaugh J.A., Patterson A.D., Pollard K.S., Mai K., Spranger J, & Turnbaugh P.J. (2021). Caloric restriction disrupts the microbiota and colonization resistance. Nature, 595(7866), 272-277.
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2

Quantifying Gut Bacterial Abundance

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Quantitative PCR (qPCR) was performed on DNA extracted from the human stool samples. DNA templates were diluted to 5 ng/µL before qPCR of total 16S rRNA gene copies was carried out in triplicate 10 µL reactions with 200 nM 340F/514R primers using a BioRad CFX384 thermocycler with SYBR Select for CFX Master Mix (Life Technologies) according to the manufacturer's instructions and an annealing temperature of 60˚C. Absolute quanti cations were determined against a standard curve of puri ed 8F/1542R ampli ed Akkermansia muciniphila DNA. Mean values of triplicate reactions were taken for further downstream analyses. Absolute bacterial abundance was derived by adjustments for dilutions during DNA extraction and template normalization dividing by the total fecal mass used for DNA extraction in grams.
Ang Q.Y., Alba D., Upadhyay V., Bisanz J.E., Cai J., Lee H.L., Barajas E., Wei G., Patterson A.D., Koliwad S.K., & Turnbaugh P.J. (2020). Differences in the Gut Microbiomes of Distinct Ethnicities Within the Same Geographic Area Are Linked to Host Metabolic Health.
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