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Epicentre masterpure kit

Manufactured by Illumina
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Sourced in United States

The Epicentre Masterpure kit is a nucleic acid extraction and purification tool designed for the isolation of high-quality genomic DNA, total RNA, or viral nucleic acids from a variety of sample types. The kit employs a rapid and efficient protocol to obtain pure, high-yield nucleic acid samples suitable for downstream applications.

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Lab products found in correlation

Steponeplus real time pcr system, by Thermo Fisher Scientific (2 mentions) High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (2 mentions) Nexteraxt library prep, by Illumina (1 mentions) Rs 2, by Pacific Biosciences (1 mentions) Dneasy blood and tissue kit, by Qiagen (1 mentions) Rnalater, by Thermo Fisher Scientific (1 mentions) Flexcycler2, by Analytik Jena (1 mentions) Mesa blue qpcr mastermix plus kit, by Eurogentec (1 mentions) Mircury lna universal rt kit, by Qiagen (1 mentions) Mx3000p real time pcr system cycler, by Agilent Technologies (1 mentions) Turbo dnase, by Thermo Fisher Scientific (1 mentions) Faststart universal sybr green master, by Roche (1 mentions)

4 protocols using epicentre masterpure kit

1

Whole-Genome Sequencing of Bacterial Isolates

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Isolates were grown on Eugon agar from frozen culture. After 48–72 hours of incubation at 37°C in 5% CO2, colonies were selected and plated on to fresh Eugon agar to ensure pure culture for extraction. DNA was extracted (Epicentre Masterpure kit, Epicentre, Madison, WI) at 48–72 hours postinoculation and sequenced using NexteraXT library prep (Illumina, San Diego, CA) on a Miseq (Illumina). For long read sequencing DNA was extracted (DNeasy Blood and Tissue kit, Qiagen, Hilden, Germany) libraries prepared with 15kb-20kb insert size (BluePippin kit) and sequenced on an RS II (Pacific Biosciences, Menlo Park, CA) with 2 SMRT cells per sample.
Hicks J., Stuber T., Lantz K., Erdman M., Robbe-Austerman S, & Huang X. (2018). Genomic diversity of Taylorella equigenitalis introduced into the United States from 1978 to 2012. PLoS ONE, 13(3), e0194253.
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2

Bombus Gut Microbiome Analysis

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Two species of Bombus (B. bimaculatus and B. impatiens) were collected at 4 locations near New Brunswick, New Jersey in spring and summer 2013, and 12 species (B. appositus, B. balteatus, B. bifarius, B. fervidus, B. flavifrons, B. frigidus, B. huntii, B. mixtus, B. nevadensis, B. occidentalis, B. rufocinctus, and B. sylvicola) were collected at 15 sites near Crested Butte, Colorado in summer 2014 (see Supplementary Table SI1 for locations, dates and additional information). Workers were captured with nets in the field and visually identified; their guts were aseptically dissected and placed in RNAlater (Ambion, Austin, TX, USA) for preservation. Preserved guts were stored at −80°C until nucleic acid was extracted. Specimen bodies were pinned and maintained as vouchers.
DNA was extracted from these samples using the Epicentre Masterpure kit (Epicentre, Madison, WI, USA) using the manufacturers’ protocol with an initial lysozyme treatment (250,000 units 1 hour at 37°C). DNA was suspended in ultrapure water and normalized to a concentration of 20 ng/μl.
Bombus species identifications were verified by amplifying, sequencing and submitting the COI gene to the Bold System V3 identification database as detailed in (Cariveau et al., 2014 (link)).
Powell J.E., Ratnayeke N, & Moran N.A. (2016). Strain diversity and host specificity in bee gut symbionts revealed by deep sampling of single copy protein-coding sequences. Molecular ecology, 25(18), 4461-4471.
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3

Quantitative Analysis of RNA Expression

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Total RNA was extracted from human kidney cells (HK2 cells) and complete UUO kidney samples using the Illumina’s Epicentre MasterPure Kit (Madison, WI, USA). Reverse transcription was performed for the miRNAs with MiRCURY LNA Universal RT Kit of Exiqon (Vedbaek, Denmark) and for the mRNA with High Capacity cDNA Reverse Transcription Kit of Thermo Scientific (Waltham, MA, USA) on a FlexCycler2 (Analytik Jena AG, Jena, Germany). Quantitative PCR amplification was performed on a StepOnePlus Real-Time PCR System (Thermo Scientific Waltham, MA, USA). Sybr Green technology was used for miRNAs according to Exiqon’s kit, while for the mRNA PCR the MESA BLUE qPCR MasterMix Plus kit from Eurogentec (Liege, Belgium). The primers used for the PCR are listed in Additional file 9.
Papadopoulos T., Casemayou A., Neau E., Breuil B., Caubet C., Calise D., Thornhill B.A., Bachvarova M., Belliere J., Chevalier R.L., Moulos P., Bachvarov D., Buffin-Meyer B., Decramer S., Auriol F.C., Bascands J.L., Schanstra J.P, & Klein J. (2017). Systems biology combining human- and animal-data miRNA and mRNA data identifies new targets in ureteropelvic junction obstruction. BMC Systems Biology, 11, 31.
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4

RNA Extraction and Quantification in Brucella

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For RNA extraction, bacteria were cultured in rich media up to an exponential growth phase. Total RNA was extracted using Epicentre ® Master Pure™ Kit (Illumina). One microgram of RNA was treated with TURBO DNase (Thermo Fisher Scientific, Waltham, MA, USA) and subjected to retro-transcription with Super Script III Reverse Transcriptase and random hexadeoxynucleotides (Thermo Fisher Scientific, Waltham, MA, USA) according to manufacturer’s instructions. cDNAs were then amplified with FastStart Universal SYBR Green Master (Roche, Basel, Switzerland) using the Mx3000P Real Time PCR System cycler (Agilent Technologies, Santa Clara, CA, USA). The results of each target were normalized to B. suis Translation Initiation Factor-1 (if-1, BR0249) mRNA (Eskra et al., 2001 (link)). The relative expression levels of the target genes were calculated according to the comparative critical threshold (ΔΔCT) method (Livak and Schmittgen, 2001 (link)). Three biological samples were measured for each strain. Oligonucleotides used are listed in Table S1.
Bialer M.G., Ferrero M.C., Delpino M.V., Ruiz-Ranwez V., Posadas D.M., Baldi P.C, & Zorreguieta A. (2021). Adhesive Functions or Pseudogenization of Type Va Autotransporters in Brucella Species. Frontiers in Cellular and Infection Microbiology, 11, 607610.
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