Molecular biology grade water

Manufactured by Quality Biological

Sourced in United States

Molecular biology grade water is a high-purity water specifically designed for use in molecular biology applications. It is free from contaminants, RNase, DNase, and pyrogens, ensuring optimal performance in sensitive procedures such as PCR, DNA sequencing, and RNA analysis.

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Lab products found in correlation

Qiaquick pcr purification kit, by Qiagen (2 mentions) Fusion tribrid mass spectrometer, by Thermo Fisher Scientific (1 mentions) Rituxan, by Genentech (1 mentions) Protein deconvolution 2, by Thermo Fisher Scientific (1 mentions) Reditux, by Roche (1 mentions) Ristova, by Roche (1 mentions) Remicade, by Johnson & Johnson (1 mentions) Phenol chloroform isoamyl alcohol, by Thermo Fisher Scientific (1 mentions) Glycoblue coprecipitant, by Thermo Fisher Scientific (1 mentions) Vacufuge plus, by Eppendorf (1 mentions) T4 polynucleotide kinase, by New England Biolabs (1 mentions) Infinity kit, by Thermo Fisher Scientific (1 mentions) Pierce bca assay, by Thermo Fisher Scientific (1 mentions)

5 protocols using molecular biology grade water

Characterization of Monoclonal Antibody Drugs

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The formulated rituximab drug product vial, Rituxan^® (Genentech), contains a protein concentration of 10 mg/mL in a 10 mL injection vial. The formulated infliximab drug product vial, Remicade ^® (Janssen), contains 100 mg of lyophilized powder for reconstitution at 10 mg/mL; in our case by adding 10 mL molecular biology grade water (Quality Biological, Gaithersburg, MD). Roughly 20 mg protein per lot was used to obtain one complete dataset from the SEC-FPLC, SEC-HPLC-MALS, 1D ¹H NMR and CD assays. The amount of protein needed can be lowered significantly if analytical SEC-FPLC and an NMR cryogenic probe are used instead of the instruments applied here. In addition, the two non-US FDA approved Indian-sourced rituximab drug product vials, Reditux^® (Dr. Reddy’s) and Ristova^® (Roche), containing mAb concentrations of 10 mg/mL in a 10 mL injection vial were used for NMR data comparison only.
For intact mass spectrometry (MS), around 0.3 mg of the mAbs were buffer exchanged from formulation into an ElectroSpray Ionization (ESI) compatible buffer (40% acetonitrile and 1% formic acid in water). Intact MS data were collected using a Fusion Tribrid Mass Spectrometer (Thermo Scientific, Bremen, Germany). The molecular weight was determined using the ReSpect algorithm within Protein Deconvolution 2.0 (Thermo Scientific). The detail experimental procedure is in the supporting information.

Chen K., Long D.S., Lute S.C., Levy M.J., Brorson K.A, & Keire D.A. (2016). Simple NMR methods for evaluating higher order structures of monoclonal antibody therapeutics with quinary structure. Journal of pharmaceutical and biomedical analysis, 128, 398-407.

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16S rRNA Gene Amplification and Illumina Sequencing

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All samples were subjected to PCR amplification [33 (link)] using universal bacterial/archaeal primer set 515f/806r, which targets V4 region of 16S rRNA gene [34 (link)]. Sample preparation followed the Earth Microbiome Project 16S rRNA Amplification Protocol [35 ]. Minor changes included using Molecular Biology Grade Water (Quality Biological, Gaithersburg, MD), and QIAquick PCR Purification Kit (QIAGEN, Valencia, CA). Pooled samples were submitted to the Virginia Bioinformatics Institute for paired-end 250 cycle Illumina sequencing (VBI GRL MiSeq sequencing service, Blacksburg, VA). Eight out of 300 samples were precluded due to low yield in PCR products. Field blank, filter blank and tube blank samples were pooled on a “maximum volume” basis instead of equal molar criteria (maximum volume of other samples in the same lane). During the preparation process, all samples experienced identical freeze-thaw cycles.

Ji P., Parks J., Edwards M.A, & Pruden A. (2015). Impact of Water Chemistry, Pipe Material and Stagnation on the Building Plumbing Microbiome. PLoS ONE, 10(10), e0141087.

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Biofilm Microbial Community Profiling by 16S Amplicon Sequencing

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Biofilm samples were subjected to PCR amplification using the universal bacterial/archaeal primer set F515 (5′ GTG CCA GCM GCC GCG GTA A 3′)/R806 (5′ GGA CTA CHV GGG TWT CTA AT 3′, barcoded, #0–95) targeting the V4 region of 16S rRNA gene, which was determined to yield optimal community clustering with this particular read length (Caporaso et al., 2011). Samples were prepared following the Earth Microbiome Project 16S rRNA Amplification Protocol version 4_13 (ftp://ftp.metagenomics.anl.gov/data/misc/EMP/SupplementaryFile1_barcoded_primers_515F_806R.txt) with the exceptions of using molecular biology grade water (Quality Biological, Gaithersburg, MD, USA) and the QIAquick PCR Purification Kit (Qiagen, Valencia, CA, USA). Of the 96 biofilm samples, 90 were pooled on an equal molar basis (200 ng), and the remaining six samples with low‐yield PCR products were pooled based on ‘maximum volume’ criteria (maximum volume of the 90 samples, which is 50 μl). Paired‐end 250‐cycle Illumina sequencing was performed by the Genomics Research Laboratory at the Virginia Bioinformatics Institute (Blacksburg, VA, USA).

Buse H.Y., Ji P., Gomez‐Alvarez V., Pruden A., Edwards M.A, & Ashbolt N.J. (2017). Effect of temperature and colonization of Legionella pneumophila and Vermamoeba vermiformis on bacterial community composition of copper drinking water biofilms. Microbial Biotechnology, 10(4), 773-788.

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Radiolabeling and Ethanol Precipitation of RNA

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The 8-oxoG containing oligomer and the unmodified oligomer were radiolabeled using T4 polynucleotide kinase (NEB, Ipswich, MA) as described by the manufacturer. After labeling, RNA was cleaned up by ethanol precipitation. This was done by first adding 1 M Tris buffer (pH 8.0) and 1 M sodium acetate (pH 5.2) to the reaction mixture to bring the final concentrations to 50 mM and 0.3 M respectively. Two volumes of phenol/chloroform/isoamyl alcohol (25:24:1) (Fisher Scientific, Hampton, NH) were then added and the solution was vortexed for one minute followed by centrifugation at 15,000 g for 2 min to achieve phase separation. The aqueous (top) phase was collected, and 1 µl of GlycoBlue Coprecipitant (Thermo Fisher, Waltham, MA) and 2.5 volumes of chilled 100% absolute ethanol (OmniPur, 200 Proof, Millipore Sigma, Burlington, MA) were added. The solution was mixed and then incubated overnight at −20 °C. The following day, the solution was centrifuged at 4 °C at 15,000 g for 15 min. The supernatant was removed and then washed with 95% ethanol followed by centrifugation at 15,000 g for 5 min. The supernatant was discarded, and the pellet was dried in a vacufuge plus (Eppendorf, Hamburg, Germany) for 5 min before resuspension in Molecular Biology Grade Water (Quality Biological, Gaithersburg, MD).

Gonzalez-Rivera J.C., Orr A.A., Engels S.M., Jakubowski J.M., Sherman M.W., O'Connor K.N., Matteson T., Woodcock B.C., Contreras L.M, & Tamamis P. (2019). Computational evolution of an RNA-binding protein towards enhanced oxidized-RNA binding. Computational and Structural Biotechnology Journal, 18, 137-152.

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Liver Lipid Quantification in Pcx Mice

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Livers harvested from fed and 24 h fasted Pcx^L−/− and Pcx^f/f mice (n = 6/biological replicates per genotype) were collected, flash frozen in liquid nitrogen, and stored at −80 °C. About 100 mg of frozen liver tissue was homogenized in 500 μl of ice-cold molecular biology grade water (Quality Biological) as previously described (36 (link)). Next, 200 μl of the liver homogenate was collected for lipid extraction with 1 ml of chloroform:methanol (2:1) and centrifuged at 300g for 5 min at 4 °C. The lower chloroform phase was collected into a new microcentrifuge tube and dried in a vacuum. The dried lipid samples were resuspended in 50 μl of tert-butanol:MeOH:Triton-X100 (3:1:1) before determining TG content using an Infinity Kit (Thermo Fisher Scientific). Protein content of the liver homogenate was measured by using a Pierce BCA assay (Thermo Fisher Scientific) and lipid levels were normalized to total protein.

Selen E.S., Rodriguez S., Cavagnini K.S., Kim H.B., Na C.H, & Wolfgang M.J. (2022). Requirement of hepatic pyruvate carboxylase during fasting, high fat, and ketogenic diet. The Journal of Biological Chemistry, 298(12), 102648.

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