Chloroform

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Chloroform is a colorless, volatile, and dense liquid chemical compound. It is commonly used in scientific research and laboratory settings as a solvent for various organic compounds.

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Lab products found in correlation

Close spelling variants of this product

Trizol/chloroform method (12 citations) Phenol/chloroform/isoamyl alcohol (121 citations) Deuterated chloroform (CDCl3) (5 citations) Certified ACS grade chloroform (2 citations) TRIzol and chloroform mixture (2 citations) Neutral phenol–chloroform-isoamyl alcohol (4 citations) TRIzol™-Chloroform protocol (2 citations) Acid phenol:chloroform MB grade (2 citations) Acid phenol/chloroform (49 citations) ACS grade chloroform (5 citations)

747 protocols using chloroform

Bacterial RNA Isolation Protocol

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For RNA isolation, 450 μL bacterial suspension (10⁵ CFU/mL in BHI) was prepared for each experimental condition, treated with CPP-PNA conjugates as indicated, and incubated in a 2-mL reaction tube for 6 h at 37°C and 7 rpm (Rotor SB3; Stuart, Staffordshire, United Kingdom). Subsequently, five samples per condition were pooled. Bacteria were pelleted immediately, shock-frozen in liquid nitrogen, and stored at −80°C until use. Total RNA was extracted according to the protocol published by Li-Korotky et al. (46 (link)). Briefly, 30 μg polyinosinic acid (Poly I) was added to each bacterial pellet. TRIzol reagent was added to the bacteria/Poly I mix, which was subsequently disrupted in a homogenizer (Peqlab Biotechnologie GmbH, Erlangen, Germany). RNA was extracted twice with 200 μL chloroform (Thermo Fisher Scientific, Darmstadt, Germany). Following the addition of 5 μL glycogen and 500 μL isopropanol, RNA was precipitated at 13,000 rcf (relative centrifugal force) at 4°C for 15 min. Pellets were suspended in H₂O. RNA was subsequently treated with acid phenol:chloroform:isoamyl alcohol (125:24:1) (pH 4.5) (Thermo Fisher Scientific) and TURBO DNase (Thermo Fisher Scientific) according to the manufacturer’s instructions. RNA was stored at −80°C until further use.

Barkowsky G., Abt C., Pöhner I., Bieda A., Hammerschmidt S., Jacob A., Kreikemeyer B, & Patenge N. (2022). Antimicrobial Activity of Peptide-Coupled Antisense Peptide Nucleic Acids in Streptococcus pneumoniae. Microbiology Spectrum, 10(6), e00497-22.

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Preparative Chromatography of Lipid Mixtures

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Starting at the base of the column, a classic preparative chromatography
column (with solvent reservoir) was prepared with a glass wool plug, washed sand
(Fisher Scientific), silica gel (VWR International #SX0143U-1) at a
ratio of 50:1 silica mass:sample mass that had been suspended in hexane (Sigma),
and another layer of washed sand on top. The hexane was drained from the column
and the lipid mixture, resuspended in hexane, was loaded slowly on to the column
and allowed to migrate through the sand into the silica gel layer. Lipids were
eluted with the following solvent system: 1 column volume hexane (Sigma), 2
column volumes dichloromethane (Fisher Scientific), 2 column volumes of 1:1
dichloromethane:chloroform (Fischer Scientific), and then chloroform until the
desired C₂₈H₄₄O₃ had completely eluted as
monitored by TLC.

Nielson J.R., Fredrickson E.K., Waller T.C., Rendón O.Z., Schubert H.L., Lin Z., Hill C.P, & Rutter J. (2017). Sterol oxidation mediates stress-responsive Vms1 translocation to mitochondria. Molecular cell, 68(4), 673-685.e6.

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EV RNA Extraction via nanoFACS

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EV suspension volume following nanoFACS sorting was approximated and Qiazol (from the exoRNeasy kit) and chloroform (ThermoFisher Scientific, Waltham, MA, USA) volumes were adjusted according to a 15 μl:70 μl:9 μl ratio (EV suspension:Qiazol:chloroform). Qiazol was added to the tube, the tube inverted several times, and briefly vortexed to ensure thorough mixing. The solution was incubated at room temperature for 10 min. The previously determined volume of chloroform was added to the tube and the solution was vigorously shaken for 15 s. From here, the exoRNeasy protocol was followed, picking up from step 10 (3 min chloroform/phenol incubation).

Hsia T., Yekula A., Batool S.M., Rosenfeld Y.B., You D.G., Weissleder R., Lee H., Carter B.S, & Balaj L. (2022). Glioblastoma‐derived extracellular vesicle subpopulations following 5‐aminolevulinic acid treatment bear diagnostic implications. Journal of Extracellular Vesicles, 11(11), 12278.

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Chloroform Extraction and Derivatization for GC-MS Analysis

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Samples were non-derivatized and derivatized for the GC-MS analysis. For derivatized samples, the preparation prior to GC-MS analysis was performed according to Semreen [18 (link)] methodology as follows: 30 mL from cell-free supernatants were extracted three times with 90 mL of chloroform (Fisher Scientific, Santa Clara, CA, USA). The chloroform layer was dehydrated over anhydrous sodium sulphate (Fisher Scientific), filtered, and followed by evaporation using a rotatory evaporator (Buchi, Essen, Germany). The residue collected from each extract was dissolved in 500 μL of chloroform prior to GC-MS injection. In addition, 100 µL of the chloroform extract was derivatized by adding 50 μL of N-trimethylsilyl-N-methyl trifluoroacetamide and trimethylchlorosilane (MSTFA + 1% TMS) followed by incubation at 50 °C for 30 min prior to GC-MS injection.

López-Ramos J.E., Bautista E., Gutiérrez-Escobedo G., Mancilla-Montelongo G., Castaño I., González-Chávez M.M, & De Las Peñas A. (2021). Analysis of Volatile Molecules Present in the Secretome of the Fungal Pathogen Candida glabrata. Molecules, 26(13), 3881.

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Candida Metabolite Extraction and GC-MS Analysis

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The filtered supernatants collected separately from each Candida isolate was extracted with chloroform (Fisher Scientific, Santa Clara, CA, USA). The chloroform layer was dehydrated over anhydrous sodium sulphate (Fisher Scientific) followed by evaporation using rotatory evaporator (Buchi, Essen, Germany). The residue collected from each extracted culture was dissolved in 500 µL chloroform prior to GC-MS injection. Furthermore, 100 µL of the chloroform extract was derivatized by adding 50 µL of N-trimethylsilyl-N-methyl trifluoroacetamide and trimethylchlorosilane (MSTFA + 1% TMS) followed by incubation at 50 °C for 30 min prior to GC-MS analysis.

Semreen M.H., Soliman S.S., Saeed B.Q., Alqarihi A., Uppuluri P, & Ibrahim A.S. (2019). Metabolic Profiling of Candida auris, a Newly-Emerging Multi-Drug Resistant Candida Species, by GC-MS. Molecules, 24(3), 399.

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Biocatalytic Oxidation of Alkene Substrates

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1-Nonene (98%), 1-heptene (97%), styrene (99%), cyclohexene (99%) and 1-methylcyclohexene (97%), cyclohexene oxide (standard), and Chloroform-d were obtained from Sigma-Aldrich, USA. 1-Methylcyclohexene oxide (standard) was bought from Tokyo Chemical Industry, Japan. Chloroform, ethyl acetate, and toluene were purchased from Fisher Scientific, UK. Acetone, H₂O₂ (30%, w/w), and phenylacetic acid were purchased from Merck, Germany. A commercially known biocatalyst, CALB, immobilised on a macroporous acrylic resin (Novozym 435) was obtained from Novozymes Corporation, Denmark. The standards for 1-Nonene oxide and 1-heptene oxide were prepared and identified as described by Rusch Gen Klaas and Warwel [15 ]. For GC-MS analysis, ethyl acetate and Chloroform HPLC-grade were procured from Fisher Scientific, UK.

Abdulmalek E., Arumugam M., Mizan H.N., Abdul Rahman M.B., Basri M, & Salleh A.B. (2014). Chemoenzymatic Epoxidation of Alkenes and Reusability Study of the Phenylacetic Acid. The Scientific World Journal, 2014, 756418.

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Lipid Extraction and Identification by TLC

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An equivalence of 2 mg of protein for each membrane was extracted by Bligh-Dyer method (73 (link)). The chloroform layer was harvested and dried under nitrogen gas, and the GPLs were resuspended in 60 μL of chloroform (Fisher Chemical no. C607-4). TLC was performed on silica gel 60 plates (Sigma no. 1057210001). The GPLs were spotted as successive 20 μL droplets onto the origin of the TLC plate and dried. For one-dimensional TLC, a mobile phase of chloroform/methanol/acetic acid (130:50:20) was used. In case of two-dimensional TLC, a mobile phase of chloroform/methanol/water (120:50:8) was used for the first dimension, and a mobile phase of chloroform/methanol/acetic acid/water (160:24:30:8) was used for the second dimension. The GPLs were visualized by iodine vapor and identified using commercial standards CL (no. 710341), PGl (no. 840457), PE (no. 850757), and PA (no. 840857) (Avanti).

Cian M.B., Mettlach J.A., Zahn A.E., Giordano N.P., Minor K.E., McClelland M, & Dalebroux Z.D. (2022). Cardiolipin Biosynthesis Genes Are Not Required for Salmonella enterica Serovar Typhimurium Pathogenesis in C57BL/6J Mice. Microbiology Spectrum, 10(3), e02617-21.

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Organic Materials Synthesis and Purification

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For this study, SubPc (99%), mCP (99%), TPBi (99%), C545T (99%), TAPC (99%), and PTB7 (Mw > 50,000) were obtained from Lumtec Inc., C₆₀ (99%) and C₇₀ (99%) were obtained from MER Corporation, MoO₃ (99%) and BCP (98%) were obtained from Alfa Aesar, chloroform (99%), 3-mercaptopropionic acid (99%), octane (98%), oleic acid (tech. grade 90%) acetonitrile (99.5%), and methyl acetate (anhydrous) were obtained from Sigma–Aldrich, selenium dioxide (99.8%) and 1-octadecene (90%) were obtained from Acros Organics. All materials above were used as received. The NPD (99%) was synthesized by the Dow Chemical Company and purified once by temperature-gradient sublimation^{69 (link)}. Cadmium myristate was prepared on a multi-gram scale according to the method reported by Chen et al.^{70 (link)}.

Zhang T., Dement D.B., Ferry V.E, & Holmes R.J. (2019). Intrinsic measurements of exciton transport in photovoltaic cells. Nature Communications, 10, 1156.

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RNA Isolation and qPCR Protocol

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At least 1.5 Million cells growing in T75 culture flasks were deprived of medium and treated with 1 ml of TRIZol (Thermofisher). Further workup occurred as per manufacturer description, using 400 µl of chloroform (Alfa Aesar, Molecular Biology Reagent, stabilized with 50 ppm amylene), Iso-Propanol (Sigma, HPLC grade) and Ethanol (Roth, p.A.). The resulting RNA was characterized with a colibri-nanodrop device from Titertek Berthold and used for cDNA synthesis using the RevertAid kit (Thermofisher). For the Q-PCR runs on a StepOnePlus instrument from Applied Biosystems, 100 ng of the resulting cDNA were used per sample using vinculin as a housekeeping gene. The resulting probe volume was 25 µl, using the Quantitect primer system from QIAGEN and evaGreen 5x Q-PCR Mastermix with ROX dye from Solis Biodyne. PCR plates were from Saarstedt. The sample data was evaluated according to the Ct method with the StepOne Software version 2.3.

Reppingen N., Helm A., Doleschal L., Durante M, & Fournier C. (2021). A Combination of Cabozantinib and Radiation Does Not Lead to an Improved Growth Control of Tumors in a Preclinical 4T1 Breast Cancer Model. Frontiers in Oncology, 11, 788182.

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Preparation of PLGA-Based Drug Delivery System

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Dexamethasone (DEX > 99%, Tokyo Chemical Industry Co., Ltd., Tokyo, Japan), beta-cyclodextrin (β-CD, Sigma-Aldrich, St. Louis, MO, USA), chloroform (> 99.8%, Alfa Aesar, Ward Hill, MA, USA), methanol (Thermo Fisher Scientific Inc., Hampton, NH, USA), 1,1,1,3,3,3 hexafluoroisopropanol (HFIP, 99%, Beantown Chemical Co., Hudson, NH, USA), Hexamethyldisilazane (> 99%, Sigma-Aldrich), hexcetylpyridinium chloride monohydrate (CPC, Sigma-Aldrich), β-glycerophosphate (Sigma-Aldrich), ascorbic acid (Sigma-Aldrich), paraformaldehyde (PFA, Sigma-Aldrich) and 75:25 poly(DL-lactide-co-glycolide acid) (PLGA, inherent viscosity: 0.55–0.75 dL/g in CHCl₃, Lactel Absorbable Polymers, Durect Corporation, Birmingham, AL, USA) were purchased and used as-received. Distilled-deionized (DI) water from a Millipore Milli-Q ultrapure water system was used in the experiments.

Daghrery A., Aytac Z., Dubey N., Mei L., Schwendeman A, & Bottino M.C. (2020). Electrospinning of dexamethasone/cyclodextrin inclusion complex polymer fibers for dental pulp therapy. Colloids and surfaces. B, Biointerfaces, 191, 111011.

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