neb cutsmart buffer (New England Biolabs)

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Name:

NEBuffer Set 1 1 2 1 3 1 and CutSmart

Description:

NEBuffer Set 1 1 2 1 3 1 and CutSmart 6 0 ml

Catalog Number:

B7200S

Price:

Category:

Buffers

Size:

6 0 ml

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New England Biolabs neb cutsmart buffer

NEBuffer Set 1 1 2 1 3 1 and CutSmart 6 0 ml
https://www.bioz.com/result/neb cutsmart buffer/product/New England Biolabs
Average 95 stars, based on 1 article reviews
Price from $9.99 to $1999.99

neb cutsmart buffer - by Bioz Stars, 2021-06

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Incubation:

Article Title: Biochemically diverse CRISPR-Cas9 orthologs
Article Snippet: Purified Cas9 protein was then diluted to 1μM in dilution buffer (300mM NaCl, 20mM Tris, pH7.5) and stored on ice. .. Cas9 and sgRNA were then combined in a 2:1 sgRNA:Cas9 molar ratio in reaction buffer at room temperature for 10 min. Substrate was added next at a Cas9:sgRNA:DNA ratio of 10:20:1 and incubated for 30 min. For buffer optimization and spacer length preference experiments, 1X NEBuffers 1.1, 2.1, 3.1, or CutSmart (NEB B7200S) were used as reaction buffers and incubations took place at 37°C. .. Cas9 and sgRNA were then combined in a 2:1 sgRNA:Cas9 molar ratio in reaction buffer at room temperature for 10 min. Substrate was added next at a Cas9:sgRNA:DNA ratio of 10:20:1 and incubated for 30 min. For buffer optimization and spacer length preference experiments, 1X NEBuffers 1.1, 2.1, 3.1, or CutSmart (NEB B7200S) were used as reaction buffers and incubations took place at 37°C.

Article Title: Biochemically diverse CRISPR-Cas9 orthologs
Article Snippet: Following purification with a Monarch® PCR & DNA Cleanup Kit and elution into 1X. .. CutSmart® buffer (NEB) containing 1 mM ATP, 15 units of Exonuclease V (RecBCD; NEB) and T5 exonuclease (NEB) were added to the sample and incubated at 37°C for 45 min. 0.04 units of Proteinase K (NEB) was then added and the sample was incubated at 25°C for 15 min. prior to purification with a Monarch® PCR & DNA Cleanup Kit. .. After elution, the yield of circular double-stranded DNA was assessed using an Agilent 2100 Bioanalyzer.

Article Title: A cell-based screening system for RNA polymerase I inhibitors cell-based screening system for RNA polymerase I inhibitors †Electronic supplementary information (ESI) available: Growth inhibition, plasmid maps and characterization. See DOI: 10.1039/c9md00227h
Article Snippet: The vector plasmid was YIPlac211-TG1, which is a yeast integrative plasmid (Fig. S6b ). .. Both plasmids were incubated in 10 μL of 1 × NEB CutSmart Buffer with two restriction endonucleases EcoRI-HF and Hind III-HF at a dose of 10 U μL–1 at 37 °C for 12 h to cut the plasmid at the corresponding loci and at 65 °C for 1 h to denature the enzyme. .. The resulting DNA segments from both reactions were separated on a 0.7% agarose gel suffused in 1 × TAE buffer with ethidium bromide using BioRad electrophoresis gel apparatus with an electric field strength of 5 volts per cm, as depicted in .

Article Title: A catalogue of biochemically diverse CRISPR-Cas9 orthologs
Article Snippet: Cas9 and sgRNA were then combined in a 2:1 sgRNA:Cas9 molar ratio in reaction buffer at room temperature for 10 min. .. The substrate was added next at a Cas9:sgRNA:DNA ratio of 10:20:1 and incubated for 30 min. For buffer optimization and spacer length preference experiments, 1× NEBuffers 1.1, 2.1, 3.1, or CutSmart (NEB B7200S) were used as reaction buffers, and incubations took place at 37 °C. ..

other:

Article Title: Transcriptome-Wide Mapping 5-Methylcytosine by m5C RNA Immunoprecipitation Followed by Deep Sequencing in Plant.
Article Snippet: Transcriptome-wide mapping RNA modification is crucial to understand the distribution and function of RNA modifications.

Article Title: Genome-wide analyses of chromatin interactions after the loss of Pol I, Pol II, and Pol III
Article Snippet: Then, the supernatant was discarded, and the nuclei were washed twice with 1× NEB CutSmart buffer.

Article Title: Systematic identification of cis-regulatory variants that cause gene expression differences in a yeast cross
Article Snippet: We digested 1 µg of barcoded library using 5 µL SfiI enzyme, 10 µL 10x NEB CutSmart buffer, and water to 100 µL at 50 °C for 2 hr.

Purification:

Polymerase Chain Reaction:

Plasmid Preparation:

Isolation:

Article Title: Structural and spatial chromatin features at developmental gene loci in human pluripotent stem cells
Article Snippet: Cross-linked nuclei were isolated by incubation in 550 μl of lysis buffer (10 mM Tris-HCl, pH 7.5, 10 mM NaCl, 0.2% Np-40, and 1× protease inhibitor (Roche 04 693 159 001)) on ice for 20 min and homogenization with 10 strokes × 2 by a tight pestle in a dounce homogenizer (Wheaton) on ice. .. The isolated nuclei were washed twice with 500 μl of 1× cutsmart buffer (NEB B7200S). ..