restriction enzyme digestion with bfuai (New England Biolabs)

New England Biolabs is a verified supplier

New England Biolabs manufactures this product

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

New England Biolabs restriction enzyme digestion with bfuai
Restriction Enzyme Digestion With Bfuai, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 75 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/restriction+enzyme+digestion/BfuAI/pm42021122-118-15-20
Average 95 stars, based on 75 article reviews

restriction enzyme digestion with bfuai - by Bioz Stars, 2026-10

95/100 stars

Images

Plasmid Preparation:

Article Title: A comprehensive functional atlas of ALK kinase domain variants reveals resistance landscape to ALK inhibitors
Article Snippet: .. Plasmid constructs were assembled using either Gibson assembly (NEBuilder HiFi DNA Assembly Master Mix, New England Biolabs) or conventional ligation following restriction enzyme digestion (T4 DNA Ligase, NEB), in accordance with the manufacturer’s instructions. .. To generate pLenti-PEmax-BSD and pLenti-PE7-BSD, the SpCas9 coding sequence in the LentiCas9-Blast plasmid (Addgene #52962) [ ] was replaced with PEmax or PE7 coding sequence, which were PCR-amplified from pCMV-PEmax (Addgene #174820) [ ] and pCMV-PE7 (Addgene #214812) [ ], respectively.

Article Title: Variants in the proteasome regulator PSMF1 cause a phenotypic spectrum from parkinsonism to perinatal lethality.
Article Snippet: .. Following amplification, clean-up and restriction enzyme digestion (XhoI, BamHI) of the PCR amplicon and pSPL3 exon trapping vector was performed prior to ligation between the vector-containing exons A and B of the linearized vector that was transformed into DH5α competent cells (NEB 5- alpha, New England Biolabs, Frankfurt, Germany), plated and incubated overnight. ..

Construct:

Ligation:

Article Title: Nanoscale dynamics of enhancer-promoter interactions during exit from pluripotency.
Article Snippet: .. Restriction enzyme digestion was performed with 750 U DpnII (NEB, R0543S) at 37 ◦C for 24 h. DpnII was subsequently inactivated at 65 ◦C for 20 min and proximity ligation took place in 1 × ligation buffer and 240 U T4 DNA ligase (Thermo Fisher Scientific, EL0013) at 16 ◦C for 18 h. Ligation reactions were centrifuged at room temperature and 500 × g for 15 min, the pellets were resuspended in 300 μl of Tris–EDTA buffer (TE buffer; Sigma– Aldrich, 93283) and incubated with 3 U proteinase K at 65 ◦C for 16 h, followed by an incubation with 7.5 mU of RNase (Roche, 11119915001) at 37 ◦C for 30 min. DNA was subsequently extracted from the samples by mixing with equal volume phenol-chloroform-isoamyl alcohol mixture (Sigma– Aldrich, 77617) and centrifugation at room temperature and 12 600 × g for 10 min in prespun 5Prime light Phase Lock gel tubes (VWR, 733-2477). .. The upper aqueous layer was then mixed with 30 μl sodium acetate 3M, 1 μl GlycoBlue Coprecipitant (Thermo Fisher Scientific, AM9515) and ethanol 75% v/v at −20 ◦C for 2 h. DNA was pelleted by centrifugation at 4 ◦C and 21 000 × g for 30 min, washed with cold 70% ethanol, resuspended in TE buffer and concentration was determined with Qubit TM 1 × double-stranded DNA (dsDNA) Broad Range assay (Thermo Fisher Scientific, Q33266), according to the manufacturer’s instructions.

Article Title: Nanoscale dynamics of enhancer–promoter interactions during exit from pluripotency
Article Snippet: .. Restriction enzyme digestion was performed with 750 U DpnII (NEB, R0543S) at 37°C for 24 h. DpnII was subsequently inactivated at 65°C for 20 min and proximity ligation took place in 1× ligation buffer and 240 U T4 DNA ligase (Thermo Fisher Scientific, EL0013) at 16°C for 18 h. Ligation reactions were centrifuged at room temperature and 500 × g for 15 min, the pellets were resuspended in 300 μl of Tris–EDTA buffer (TE buffer; Sigma–Aldrich, 93283) and incubated with 3 U proteinase K at 65°C for 16 h, followed by an incubation with 7.5 mU of RNase (Roche, 11119915001) at 37°C for 30 min. DNA was subsequently extracted from the samples by mixing with equal volume phenol-chloroform-isoamyl alcohol mixture (Sigma–Aldrich, 77617) and centrifugation at room temperature and 12 600 × g for 10 min in prespun 5Prime light Phase Lock gel tubes (VWR, 733-2477). .. The upper aqueous layer was then mixed with 30 μl sodium acetate 3M, 1 μl GlycoBlue Coprecipitant (Thermo Fisher Scientific, AM9515) and ethanol 75% v/v at −20°C for 2 h. DNA was pelleted by centrifugation at 4°C and 21 000 × g for 30 min, washed with cold 70% ethanol, resuspended in TE buffer and concentration was determined with Qubit TM 1× double-stranded DNA (dsDNA) Broad Range assay (Thermo Fisher Scientific, Q33266 ), according to the manufacturer’s instructions.

Amplification:

Article Title: Mast cell-specific CysLT2 receptor signaling inhibits cysteinyl leukotriene-dependent mast cell activation and type 2 allergic lung inflammation
Article Snippet: Cloning vectors (pUCIDT-Amp) carrying variants of CysLT 2 R receptors (hM201V and hL129Q ) were purchased from Integrated DNA Technologies (IDT). .. The synthetic gene sequences were amplified by PCR (Phusion, New England Biolabs) using primers containing restriction enzyme recognition sequences and the gene fragments were subcloned into the expression vectors: pEF1α-Myc (N-terminal Myc, Clontech, Cat#631991) after restriction enzyme digestion with MSCI and NotI (New England Biolabs). .. The Coding DNA Sequences for human AC2,4,5,6,7, and 9 in pcDNA3.1 (GenScript) were amplified by PCR and the gene fragments were subcloned into the NanoBiT expression vectors: N or C-terminal Large BiT (LgBiT) and further pEF4 Vector or pF4ACMV Vector for better expression after Restriction enzyme digestion with Kpn1 and PmeI (New England Biolabs).

Article Title: Mast cell-specific CysLT2 receptor signaling inhibits cysteinyl leukotriene-dependent mast cell activation and type 2 allergic lung inflammation.
Article Snippet: For MC degranulation assay, human CBMCs were sensitized overnight using human myeloma IgE (2 μg/ mL), then stimulated with anti-IgE (1 μg/mL) for 30 min. Subcloning of the CysLT2R variants, NanoBiT constructs, Gβγ scavenger Cloning vectors (pUCIDT-Amp) carrying variants of CysLT2R receptors (hM201V 58 and hL129Q59) were purchased from Integrated DNA Technologies (IDT). .. The synthetic gene sequences were amplified by PCR (Phusion, New England Biolabs) using primers containing restriction enzyme recognition sequences and the gene fragments were subcloned into the expression vectors: pEF1α-Myc (N-terminal Myc, Clontech, Cat#631991) after restriction enzyme digestion with MSCI and NotI (New England Biolabs). .. The Coding DNA Sequences for human AC2,4,5,6,7, and 9 in pcDNA3.1 (GenScript) were amplified by PCR and the gene fragments were subcloned into the NanoBiT expression vectors: N or C-terminal Large BiT (LgBiT) and further pEF4 Vector or pF4ACMV Vector for better expression after Restriction enzyme digestion with Kpn1 and PmeI (New England Biolabs).

Polymerase Chain Reaction:

Article Title: Large-scale discovery of neural enhancers for cis-regulation therapies
Article Snippet: DNA was eluted in 50 uL EB and then a 2nd PCR reaction was performed with 5BC-AG-f02 and 5BC-AG-r02 primers to add 15 bp transcribed MPRA barcodes using Kapa HiFi Hotstart readymix (KAPA Biosystems, Cat. No. 08202940001; cycling conditions: 98°C for 3 minutes, 10 cycles of 98°C x 20 seconds, 65°C x 15 seconds, and 72°C x 15 seconds, and 72°C x 1 minute). .. The PCR product was combined in a DNA low bind tube and purified using 0.75x SPRI beads. pLS-SceI backbone vector (Addgene, Cat. No. 137725) was linearized by restriction enzyme digestion with AgeI-HF (NEB, Cat. No. R3552S) and SbfI-HF (NEB, Cat. No. R3642S) in CutSmart buffer for 1 hour at 37°C. ..

Expressing:

Incubation:

Centrifugation:

Transformation Assay:

Purification:

Article Title: Nucleoplasmic Lamin A/C controls replication fork restart upon stress by modulating local H3K9me3 and ADP-ribosylation levels.
Article Snippet: .. Restriction enzyme digestion followed (120 U of PvuII HF, New England Biolabs) in order to digest 6 μg of the purified genomic DNA for 5 h at 37oC. .. RNase A (Sigma–Aldrich, R5503) to a final concentration of 250 ug/ml was added for the last 2 h of this incubation.

Review