Perfluoro 1 octanol

Manufactured by Merck Group

Sourced in Germany

Perfluoro-1-octanol is a fluorinated organic compound with the chemical formula C8F17OH. It is a colorless liquid at room temperature with a high boiling point and low surface tension. The product is used in various industrial and laboratory applications as a specialty chemical.

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Lab products found in correlation

Kapa hifi hotstart readymix, by Roche (3 mentions) Hfe 7500, by Microfluidics (2 mentions) Exo resistant random primer, by Thermo Fisher Scientific (1 mentions) Quant it picogreen dsdna assay, by Thermo Fisher Scientific (1 mentions) Tris hcl, by Merck Group (1 mentions) A5030, by Merck Group (1 mentions) Sybr green 1, by Thermo Fisher Scientific (1 mentions) Isopropanol, by Merck Group (1 mentions) Kapa2g robust hotstart readymix, by Roche (1 mentions) Q5 high fidelity 2x master mix, by New England Biolabs (1 mentions) Vortex genie 2, by Scientific Industries (1 mentions) Su 8 3025, by MicroChem (1 mentions) Syringe pump, by Harvard Apparatus (1 mentions) Pico surf 1, by Microfluidics (1 mentions) Sarkosyl, by Merck Group (1 mentions) 75 cycle high output kit v2, by Illumina (1 mentions) Dithiothreitol (dtt), by Thermo Fisher Scientific (1 mentions) Maxima h minus reverse transcriptase, by Thermo Fisher Scientific (1 mentions) Spriselect beads, by Beckman Coulter (1 mentions) Nextseq 500, by Illumina (1 mentions)

Spelling variants of this product

1-octanol (141 citations) Octanol (39 citations) 2H-perfluoro-1-octanol (4 citations) -Perfluoro-1-octanol (PFO; (2 citations)

14 protocols using perfluoro 1 octanol

Whole Genome Amplification by MDA

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DNA was denatured by mixing the DNA diluted in milliQ water 1:1 with 50 mM KOH (Sigma Aldrich) and incubating for 3 min at room temperature (RT). The denatured DNA was the neutralized by adding an equal volume of Tris-HCl (80 mM, pH4; Sigma Aldrich). RepliPHI Phi29 Reagent Kit (Epicenter) supplemented with Exo-Resistant Random Primer (ThermoFisher Scientific) was used for the MDA reaction. A 2× MDA mastermix (2× reaction buffer, 2 mM dNTP, 50 μM primer, 4 U/μl Phi29, 8 mM DTT and 5 % DMSO) was prepared. The denatured and neutralized DNA and the 2× MDA mastermix were mixed at equal volumes by pipetting for a bulk reaction in tube or in the microfluidic chip as described above for emulsion generation. Reactions were incubated for 12 h at 30 °C. The polymerase was then inactivated at 65 °C for 10 min.
After incubation, the emulsion was broken by adding 5 μl 1H, 1H, 2H, 2H, Perfluoro-1-octanol (Sigma Aldrich), vortexing, and centrifuging briefly until the emulsion separated into one aqueous and one oil phase. If the emulsion did not break, the emulsion breaking procedure was repeated. The supernatant (aqueous phase) was collected by pipetting and could then be treated like the MDA products from the bulk reactions. The concentrations of MDA products were quantified with Qubit dsDNA kit (ThermoFisher Scientific) or Quant-iT PicoGreen dsDNA assay (ThermoFisher Scientific).

Hammond M., Homa F., Andersson-Svahn H., Ettema T.J, & Joensson H.N. (2016). Picodroplet partitioned whole genome amplification of low biomass samples preserves genomic diversity for metagenomic analysis. Microbiome, 4, 52.

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Microfluidic Droplet Generation for Single-Cell Encapsulation

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A microfluidic device with cross junction was fabricated as previously reported [49 (link)] for generating monodispersed picoliter-sized droplets. Ultra-low gelling temperature agarose A5030 (Sigma-Aldrich) was mixed with DPBS (3% w/v) and incubated at 85 °C for 30 min. The cell concentration was calculated with bacterial counter after SYBR Green I (S7563, Thermo Fisher Scientific) staining, and mixed with agarose solutions to prepare 1.5% w/v agarose cell suspensions with 3.0 × 10³ cells/μL. Agarose cell suspensions were loaded into a PTFE tube (AWG 24) connected to a Mitos P-pump (Dolomite, Charleston, MA, USA) and introduced into a microfluidic device with 2% Pico-Surf1 in Novec7500 (Dolomite) as carrier oil. Microfluidic droplets with a diameter of 40 μm (volume: 34 pL) were generated for encapsulation of >100,000 single cells within 30 min (35,000 droplets/min) at a concentration of 0.1 cell/droplet. Droplets were collected in 1.5 mL tubes from the outlet, and incubated on ice for 15 min, and broken by 1H,1H,2H,2H-perfluoro-1-octanol (Sigma-Aldrich). Gel beads were washed sequentially with 500 μL of acetone (Sigma-Aldrich), isopropanol (Sigma-Aldrich), and DPBS (DPBS for three times).

Nishikawa Y., Kogawa M., Hosokawa M., Wagatsuma R., Mineta K., Takahashi K., Ide K., Yura K., Behzad H., Gojobori T, & Takeyama H. (2022). Validation of the application of gel beads-based single-cell genome sequencing platform to soil and seawater. ISME Communications, 2, 92.

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Bead-based Bulk Suppression PCR Protocol

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Loaded beads were mixed with a polymerase master mix, KAPA2G Robust HotStart ReadyMix (KAPA Biosystems, Wilmington, MA), KAPA HiFi HotStart ReadyMix (KAPA Biosystems) or Q5 High-Fidelity 2X Master Mix (New England Biolabs), 60-nt primer sequences containing 20-nt amplification primer sequences and 40-nt inverted terminal repeats (ITRs) to be used during bulk suppression PCR (Supplementary Table S5), bovine serum albumin and BtsI-v2. Immediately after adding BtsI-v2, the mixture was added to 600 μl of BioRad Droplet Generation Oil and vortexed for 3 min using a Vortex Genie 2 (Scientific Industries, Bohemia, NY), resulting in compartmentalization of beads in <5 μm droplets. After vortexing, samples were aliquoted into PCR strips and incubated at 55°C for 90 min, allowing BtsI-v2 to cleave oligo sequences off the beads. Samples were heated to 94°C for 2 min, and then thermocycled for 60 cycles with the following conditions: 94°C for 15 s, 57°C for 20 s, 72°C for 45 s, followed by a final 5 min extension at 72°C. Following assembly, emulsions were broken by adding 100 μl perfluoro-1-octanol (Sigma-Aldrich, St Louis, MO), and the aqueous phase was extracted and column cleaned. Assembled products were then run on a 2% agarose gel and bands were extracted at the correct assembly length.

Sidore A.M., Plesa C., Samson J.A., Lubock N.B, & Kosuri S. (2020). DropSynth 2.0: high-fidelity multiplexed gene synthesis in emulsions. Nucleic Acids Research, 48(16), e95.

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Droplet Microfluidics for Malaria Detection

Cited 2 times

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Unless otherwise stated in the result section pTopI was extracted from blood or saliva from malaria patients using pump driven droplet microfluidics. The droplet microfluidic devices (see Fig. 5A for design) were fabricated by conventional soft lithography techniques^{59 (link)}, casting and curing the PDMS prepolymer on a SU-8 3025 (MicroChem) master of a channel height at around 25 μm. PDMS prepolymer (Sylgard 184) was prepared in a 10:1 (base:curing agent) ratio and cured at 65 °C for 1 hour. Prior to the experiments, the channels were wetted with oil/surfactant (Pico-Surf 1, 2% in HFE-7500, Dolomite Microfluidics) for at least 15 minutes. Two syringe pumps (Harvard Apparatus) were used to control the flow rates of oil/surfactant and reagents independently. The droplet volume and generation frequency were controlled by the flow rate ratio, determined by the competition between continuous phase and disperse phase^{34 (link),60 (link),61 (link)}. Blood/saliva (diluted 1:4 in PBS), S1 substrate (Table 1 and¹² − 167 nM final concentration in droplets) and lysis buffer (10 mM Tris pH 7.5; 5 mM EDTA; 0.2% Tween 20; 1 mM DTT; 1 mM PMSF) were subjected to droplet microfluidics essentially as described previously^{34 (link)}. The procedure was completed by breaking the droplets by addition of 25%(v/v) 1 H,1 H,2 H,2H-Perfluoro-1-octanol (Sigma-Aldrich).

Hede M.S., Fjelstrup S., Lötsch F., Zoleko R.M., Klicpera A., Groger M., Mischlinger J., Endame L., Veletzky L., Neher R., Simonsen A.K., Petersen E., Mombo-Ngoma G., Stougaard M., Ho Y.P., Labouriau R., Ramharter M, & Knudsen B.R. (2018). Detection of the Malaria causing Plasmodium Parasite in Saliva from Infected Patients using Topoisomerase I Activity as a Biomarker. Scientific Reports, 8, 4122.

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Shaken Emulsion Formation for MDA Reactions

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Shaken emulsions are generated by adding 30 μl of HFE-7500 fluorinated oil (3M, catalog no. 98-0212-2928-5) and 2% (w/w) PEG-PFPE amphiphilic block copolymer surfactant (RAN Technologies, catalog no. 008- FluoroSurfactant-1G) to 30 μl of MDA reaction mixture. Alternatively, HFE-7500 fluorinated oil with 2% PicoSurf1 (Dolomite Microfluidics) can be used. The combined mixture is vortexed at 3000 rpm for 10 s using a VWR vortexer, creating droplets ranging in diameter from 15 μm to 250 μm (Supplementary Figure S1). At the conclusion of incubation, 10 μl of perfluoro-1-octanol (Sigma Aldrich) is added, the mixture vortexed to coalesce the droplets and the aqueous layer extracted with a pipette. A detailed protocol for shaken emulsion formation can be found in Supplementary Protocol S1.

Sidore A.M., Lan F., Lim S.W, & Abate A.R. (2015). Enhanced sequencing coverage with digital droplet multiple displacement amplification. Nucleic Acids Research, 44(7), e66.

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sNucDrop-seq for Profiling Cortical Nuclei

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sNucDrop-seq of cortical nuclei was performed as previously described^{36 (link)}. Briefly, nuclei suspensions were run through an Aquapel-coated PDMS microfluidic device (uFluidix) with barcoded beads (ChemGenes) to co-encapsulate individual nuclei with a single bead. barcoded beads were resuspended in lysis buffer (200 mM Tris-HCl pH 8.0, 20 mM EDTA, 6% Ficoll PM-400 (GE Healthcare/Fisher Scientific), 0.2% Sarkosyl (Sigma-Aldrich), and 50 mM DTT (Fermentas) at a concentration of 120 beads/uL. Droplet breakage with Perfluoro-1-octanol (Sigma-Aldrich), reverse transcription using Maxima H Minus Reverse Transcriptase (ThermoFisher) and exonuclease I treatment were subsequently performed. cDNA was amplified by PCR (KAPA HiFi hotstart Readymix, KAPA biosystems) using a pre-determined, optimized number of cycles and purified twice with 0.6X SPRISelect beads (Beckman Coulter). cDNA was then tagmented using the Nextera XT DNA sample preparation kit (Illumina, cat# FC-131-1096) and further amplified using 12 enrichment PCR cycles. Libraries were sequenced on an Illumina NextSeq 500 using the 75-cycle High Output v2 Kit (Illumina), each loaded at a concentration of 2.0 pM. In total, 8 individual mouse cortex samples (4 controls, 4 CUS) were analyzed by sNucDrop-seq with 2 independent batches (2 controls, 2 CUS samples per batch), which were sequenced twice.

Kwon D.Y., Xu B., Hu P., Zhao Y.T., Beagan J.A., Nofziger J.H., Cui Y., Phillips-Cremins J.E., Blendy J.A., Wu H, & Zhou Z. (2022). Neuronal Yin Yang1 in the prefrontal cortex regulates transcriptional and behavioral responses to chronic stress in mice. Nature Communications, 13, 55.

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Encapsulation of Yeast Cells in Agarose Microgels

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The yeast culture was washed with PBS 2 times before being resuspended in PBS at an appropriate concentration based on hemocytometer counting. The low melting agarose (Sigma) solution was made with heating 2% ultra low melting agarose in PBS at 90 °C until completely melted. The melted agarose was quickly loaded in a syringe and installed to the pump. A tabletop space heater set to 80 °C was positioned to keep the agarose syringe warm during droplet generation. HFE7500 oil with 2% ionic krytox as surfactant was used for the oil phase. The agarose droplets were collected into a 50 ml falcon tube placed on ice for the formation of agarose microgel. The agarose microgel were released from the droplets by adding 20% PFO (1H,1H,2H,2H-Perfluoro-1-octanol, sigma) in HFE7500 into the emulsion followed by washing twice with TETW solution (10mM Tris pH 8.0, 1mM EDTA, 0.01% Tween-20). The agarose microgels were then resuspended in appropriate media for overnight culturing to form single colony containing microgels.

Liu L., Dalal C., Heineike B, & Abate A. (2019). High throughput gene expression profiling of yeast colonies with microgel-culture Drop-seq. Lab on a chip, 19(10), 1838-1849.

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Pressure-dependent Electric Field Droplet Release

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To determine the efficiency of the pressure-dependent
release rates by electric fields compared to the destabilizing surfactant
(1H,1H,2H,2H-perfluoro-1-octanol, Sigma-Aldrich, Germany), the droplets
were injected (pneumatic flow controller MFCS-EZ, Fluigent, Germany)
at different pressure rates into the microfluidic release device (Figure S1B,C). The applied pressures in the experiments
ranged from 50 to 300 mbar for the different inlet channels and were
adjusted as follows (for tested conditions, see Table S1). First, to get nicely separated droplets, the inlets
of the droplet channel and the separation channel were set to 50 and
45 mbar, respectively. Next, the continuous aqueous phase inlet pressure
was adjusted to 70 mbar to get a stable-phase interface in the release
channel. To release the content of the droplets into the continuous
aqueous phase, an electric field of 800 V at 1 kHz was applied to
the electrodes of the microfluidic device. Under these settings, the
release of the droplets was very successful even for higher passing
frequencies. For the release with a destabilizing surfactant, the
separation liquid was exchanged from the pure HFE 7500 oil to a pure
destabilizing surfactant solution. To obtain statistics on the droplet
release rate and the release frequency, three high-speed camera videos
of each condition were recorded and analyzed (Videos S1–S4).

Frey C., Göpfrich K., Pashapour S., Platzman I, & Spatz J.P. (2020). Electrocoalescence of Water-in-Oil Droplets with a Continuous Aqueous Phase: Implementation of Controlled Content Release. ACS Omega, 5(13), 7529-7536.

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Emulsion Formation and Disruption for Cell Recovery

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Emulsions were produced by mixing 150 ul of culture with 350 ul of FC-40 oil containing 2% (w/v) 008-FluoroSurfactant (Ran Biotechnologies) in centrifuge tubes (VWR 76332–074). Note that emulsion stability is influenced by the type of centrifuge tube used. The tubes were then gently tapped before being vortexed at max speed for 30 sec. The tubes were allowed to rest before being opened and sealed with parafilm for the 48h incubation time. To break the emulsion, the tubes were first spun down with a counter top centrifuge, for 30 sec, the lower oil phase was removed before the addition of 100 ul of 1H,1H,2H,1H-Perfluoro-1-octanol (Sigma Aldrich) mixed by pipetting up and down and gently turning the tube. After 10 min of rest, tubes were spun down for 30 sec on a counter top centrifuge, the lower phase was removed before gently mixing the aqueous phase and recovering the cells by pipetting.

Barrere J., Nanda P, & Murray A.W. (2023). Alternating selection for dispersal and multicellularity favors regulated life cycles. Current biology : CB, 33(9), 1809-1817.e3.

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Microgel Purification and Sterilization

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Excess oil was first removed by pipetting. The emulsion was broken by adding a solution of 20 wt% 1H,1H,2H,2H-Perfluoro-1-octanol (Sigma) in Novec 7500 Oil approximately equal to the volume of remaining microgels. Microgels were swelled and dispersed by adding HEPES buffer (25 mM, pH 7.4). The remaining oil was removed by washing in hexane 3x. For cell experiments, the microgels were then sterilized by washing 3x with 70% isopropyl alcohol. Finally, the microgels were washed with sterile HEPES buffer 3x where they were kept until use.

Bohlson S.S., Strasser J.A., Bower J.J, & Schorey J.S. (2001). Role of Complement in Mycobacterium avium Pathogenesis: In Vivo and In Vitro Analyses of the Host Response to Infection in the Absence of Complement Component C3. Infection and Immunity, 69(12), 7729-7735.

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