mfei restriction enzyme  (New England Biolabs)


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    Name:
    MfeI
    Description:
    MfeI 2 500 units
    Catalog Number:
    r0589l
    Price:
    302
    Size:
    2 500 units
    Category:
    Restriction Enzymes
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    Structured Review

    New England Biolabs mfei restriction enzyme
    MfeI
    MfeI 2 500 units
    https://www.bioz.com/result/mfei restriction enzyme/product/New England Biolabs
    Average 90 stars, based on 29 article reviews
    Price from $9.99 to $1999.99
    mfei restriction enzyme - by Bioz Stars, 2020-04
    90/100 stars

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    Related Articles

    Clone Assay:

    Article Title: CRISPR-Cas9 fusion to dominant-negative 53BP1 enhances HDR and inhibits NHEJ specifically at Cas9 target sites
    Article Snippet: For the tNGFR Cas9-DN1S construct, SpCas9.TGS.DN1S was moved from the PX458M plasmid construct into the CMV.tNGFR lentiviral construct between NcoI and MfeI (NEB). .. Clones were screened for the correct sequence, then used to generate dSpCas9 and dSpCas9-DN1S constructs without gRNA and with gRNA to target the human CD45 locus.

    Article Title: The β3‐integrin endothelial adhesome regulates microtubule‐dependent cell migration
    Article Snippet: .. Generation of human β3 integrin expressing cells 1 × 106 β3NULL endothelial cells were transfected with 10 μg of MfeI (New England Biolabs, Hitchin, UK) linearised full‐length human β3‐integrin (see Robinson et al ) cloned into pcDNA™ 6.2/C‐EmGFP (see Amaxa nucleofection above). .. An empty vector (EV) was used as a control.

    Centrifugation:

    Article Title: The Transcriptional Enhancer of the Pea Plastocyanin Gene Associates with the Nuclear Matrix and Regulates Gene Expression through Histone Acetylation
    Article Snippet: Nuclear matrices were collected by centrifugation, and both the supernatant and pellet fractions were treated with proteinase K at 1 mg/mL in 1% SDS at 65°C for 1 h. DNA was extracted from the pellet and supernatant fractions by phenol/chloroform extraction and ethanol precipitation. .. After six washes with buffer D (20 mM Tris-HCl, pH 8.0, 70 mM NaCl, 20 mM KCl, 10 mM MgCl2 , 0.125 mM spermidine, 0.05 mM spermine, and 0.1% digitonin), the matrices were suspended in 500 μL of buffer D, and 200 units of PvuII, MfeI, and SacI (New England Biolabs, Beverly, MA) were added.

    Amplification:

    Article Title: A Novel Mechanism of Resistance to Mouse Mammary Tumor Virus Infection
    Article Snippet: .. After amplification and agarose gel electrophoresis, the products were transferred to nylon membranes and hybridized with a probe specific to the MMTV LTR ( Semiquantitative RT-PCR analysis was accomplished by synthesis of cDNA from RNA isolated from viral particles with MMTV(C3H) LTR-specific primers, followed by digestion with Mfe I (New England Biolabs, Beverly, Mass.) as previously described ( ). .. After amplification and agarose gel electrophoresis, the products were transferred to nylon membranes and hybridized with a probe specific to the MMTV LTR ( ).

    Construct:

    Article Title: CRISPR-Cas9 fusion to dominant-negative 53BP1 enhances HDR and inhibits NHEJ specifically at Cas9 target sites
    Article Snippet: .. For the tNGFR Cas9-DN1S construct, SpCas9.TGS.DN1S was moved from the PX458M plasmid construct into the CMV.tNGFR lentiviral construct between NcoI and MfeI (NEB). .. Both DN1S and Cas9-DN1S tNGFR constructs were driven by the MND promoter.

    Incubation:

    Article Title: Hygromycin B and Apramycin Antibiotic Resistance Cassettes for Use in Campylobacter jejuni
    Article Snippet: A-ends were incorporated on the purified products by incubation with Taq DNA polymerase (Invitrogen) and dATP. .. The initial aph (7″) PCR product was instead digested with Mfe I and Sph I (NEB), purified, and ligated to low-copy pBAD24 digested with Eco RI and Sph I.

    Article Title: The Transcriptional Enhancer of the Pea Plastocyanin Gene Associates with the Nuclear Matrix and Regulates Gene Expression through Histone Acetylation
    Article Snippet: After six washes with buffer D (20 mM Tris-HCl, pH 8.0, 70 mM NaCl, 20 mM KCl, 10 mM MgCl2 , 0.125 mM spermidine, 0.05 mM spermine, and 0.1% digitonin), the matrices were suspended in 500 μL of buffer D, and 200 units of PvuII, MfeI, and SacI (New England Biolabs, Beverly, MA) were added. .. After 2 h, 200 units more of each enzyme was added, and the digestion mixture was incubated at 37°C for another 2 h. A 200-μL aliquot was removed as a total DNA sample.

    Article Title: Restriction Endonucleases from Invasive Neisseria gonorrhoeae Cause Double-Strand Breaks and Distort Mitosis in Epithelial Cells during Infection
    Article Snippet: Portions of lysates were further heat inactivated (HI) at 95°C for 10 min. For each reaction, 1 µg of VK2/E6E7 DNA (equivalent to 230,000 cells) was incubated with 45 µl bacterial lysates in a final volume of 50 µl for 24 h at 37°C and 5% CO2 . .. As positive controls, 0.5 µl each of the restriction enzymes NgoMIV, BamHI, KpnI and MfeI (New England Biolabs, Ipswich, MA, USA) and 5 µl CutSmart buffer were added to 1 µg genomic DNA in 50 µl reactions.

    Expressing:

    Article Title: CRISPR-Cas9 fusion to dominant-negative 53BP1 enhances HDR and inhibits NHEJ specifically at Cas9 target sites
    Article Snippet: Alternate truncated NGFR (tNGFR) vectors expressing DN1S or Cas9-DN1S were also developed. .. For the tNGFR Cas9-DN1S construct, SpCas9.TGS.DN1S was moved from the PX458M plasmid construct into the CMV.tNGFR lentiviral construct between NcoI and MfeI (NEB).

    Article Title: Hygromycin B and Apramycin Antibiotic Resistance Cassettes for Use in Campylobacter jejuni
    Article Snippet: The initial aph (7″) PCR product was instead digested with Mfe I and Sph I (NEB), purified, and ligated to low-copy pBAD24 digested with Eco RI and Sph I. .. The design of the pRRH and pRRA gene delivery and expression plasmids is also described in the text.

    Article Title: The β3‐integrin endothelial adhesome regulates microtubule‐dependent cell migration
    Article Snippet: .. Generation of human β3 integrin expressing cells 1 × 106 β3NULL endothelial cells were transfected with 10 μg of MfeI (New England Biolabs, Hitchin, UK) linearised full‐length human β3‐integrin (see Robinson et al ) cloned into pcDNA™ 6.2/C‐EmGFP (see Amaxa nucleofection above). .. An empty vector (EV) was used as a control.

    Modification:

    Article Title: Small- and Large-Effect Quantitative Trait Locus Interactions Underlie Variation in Yeast Sporulation Efficiency
    Article Snippet: Markers were identified and assayed using a modified restriction site-associated DNA (RAD) tag approach ( ). .. In 96 well plates, approximately 0.3 μg of DNA from each strain was digested with 5 units each of Mfe I and Mbo I [New England Biolabs (NEB)] in NEB buffer 4 for 1 hr at 37°, then heat inactivated for 20 min at 65°.

    Article Title: Enhanced Striatal Dopamine Transmission and Motor Performance with LRRK2 Overexpression in Mice is Eliminated by familial Parkinson's Disease Mutation G2019S
    Article Snippet: The following primers were used for generating homologous arms in BAC modification. .. The following primers were used to clone the fragments from genomic tail DNA containing the Mfe I restriction enzyme site and subsequently digested by Mfe I (New England Biolabs).

    Western Blot:

    Article Title: The β3‐integrin endothelial adhesome regulates microtubule‐dependent cell migration
    Article Snippet: Generation of human β3 integrin expressing cells 1 × 106 β3NULL endothelial cells were transfected with 10 μg of MfeI (New England Biolabs, Hitchin, UK) linearised full‐length human β3‐integrin (see Robinson et al ) cloned into pcDNA™ 6.2/C‐EmGFP (see Amaxa nucleofection above). .. Cells surviving 2 weeks were analysed for β3‐integrin expression by Western blotting.

    Transformation Assay:

    Article Title: Hygromycin B and Apramycin Antibiotic Resistance Cassettes for Use in Campylobacter jejuni
    Article Snippet: The purified products were then introduced to linearized pGEM-T Easy (Novagen), ligated overnight with T4 DNA ligase (NEB), and transformed into E. coli DH5α (Invitrogen). .. The initial aph (7″) PCR product was instead digested with Mfe I and Sph I (NEB), purified, and ligated to low-copy pBAD24 digested with Eco RI and Sph I.

    Transfection:

    Article Title: The β3‐integrin endothelial adhesome regulates microtubule‐dependent cell migration
    Article Snippet: .. Generation of human β3 integrin expressing cells 1 × 106 β3NULL endothelial cells were transfected with 10 μg of MfeI (New England Biolabs, Hitchin, UK) linearised full‐length human β3‐integrin (see Robinson et al ) cloned into pcDNA™ 6.2/C‐EmGFP (see Amaxa nucleofection above). .. An empty vector (EV) was used as a control.

    Ligation:

    Article Title: Small- and Large-Effect Quantitative Trait Locus Interactions Underlie Variation in Yeast Sporulation Efficiency
    Article Snippet: In 96 well plates, approximately 0.3 μg of DNA from each strain was digested with 5 units each of Mfe I and Mbo I [New England Biolabs (NEB)] in NEB buffer 4 for 1 hr at 37°, then heat inactivated for 20 min at 65°. .. The digested DNA from each strain to be genotyped was ligated to one pair of modified Illumina sequencing adapters (IDT, sequences available in supporting information, ) containing 1 of 48 different 4-bp barcodes using 1000 units of T4 ligase (NEB) in 1× ligation buffer containing dATP.

    Article Title: CRISPR-Cas9 fusion to dominant-negative 53BP1 enhances HDR and inhibits NHEJ specifically at Cas9 target sites
    Article Snippet: The overhangs generated by the BsmBI sites correspond to the AflIII and Acc65I overhangs, allowing for ligation of the U6-gRNA cassette ahead of the EFS promoter. .. For the tNGFR Cas9-DN1S construct, SpCas9.TGS.DN1S was moved from the PX458M plasmid construct into the CMV.tNGFR lentiviral construct between NcoI and MfeI (NEB).

    Article Title: Hygromycin B and Apramycin Antibiotic Resistance Cassettes for Use in Campylobacter jejuni
    Article Snippet: The initial aph (7″) PCR product was instead digested with Mfe I and Sph I (NEB), purified, and ligated to low-copy pBAD24 digested with Eco RI and Sph I. .. The ligation was transformed into E. coli DH5α and sequencing of the transformants indicated the correct aph (7″) sequence was incorporated.

    Introduce:

    Article Title: Enhanced Striatal Dopamine Transmission and Motor Performance with LRRK2 Overexpression in Mice is Eliminated by familial Parkinson's Disease Mutation G2019S
    Article Snippet: To generate LRRK2-G2019S mice, the already-modified mouse LRRK2-Wt BAC was used as the template to introduce mutation G2019S. .. The following primers were used to clone the fragments from genomic tail DNA containing the Mfe I restriction enzyme site and subsequently digested by Mfe I (New England Biolabs).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: A Novel Mechanism of Resistance to Mouse Mammary Tumor Virus Infection
    Article Snippet: .. After amplification and agarose gel electrophoresis, the products were transferred to nylon membranes and hybridized with a probe specific to the MMTV LTR ( Semiquantitative RT-PCR analysis was accomplished by synthesis of cDNA from RNA isolated from viral particles with MMTV(C3H) LTR-specific primers, followed by digestion with Mfe I (New England Biolabs, Beverly, Mass.) as previously described ( ). .. After amplification and agarose gel electrophoresis, the products were transferred to nylon membranes and hybridized with a probe specific to the MMTV LTR ( ).

    Article Title: Enhanced Striatal Dopamine Transmission and Motor Performance with LRRK2 Overexpression in Mice is Eliminated by familial Parkinson's Disease Mutation G2019S
    Article Snippet: The following primers were used to clone the fragments from genomic tail DNA containing the Mfe I restriction enzyme site and subsequently digested by Mfe I (New England Biolabs). .. The following primers were used to amplify the kinase domain by RT-PCR using mRNA isolated from transgenic and control brains and the cDNA was purified and digested by Mfe I .

    Generated:

    Article Title: CRISPR-Cas9 fusion to dominant-negative 53BP1 enhances HDR and inhibits NHEJ specifically at Cas9 target sites
    Article Snippet: The overhangs generated by the BsmBI sites correspond to the AflIII and Acc65I overhangs, allowing for ligation of the U6-gRNA cassette ahead of the EFS promoter. .. For the tNGFR Cas9-DN1S construct, SpCas9.TGS.DN1S was moved from the PX458M plasmid construct into the CMV.tNGFR lentiviral construct between NcoI and MfeI (NEB).

    Article Title: Enhanced Striatal Dopamine Transmission and Motor Performance with LRRK2 Overexpression in Mice is Eliminated by familial Parkinson's Disease Mutation G2019S
    Article Snippet: LRRK2-Wt mice were generated using the BAC technique as previously ( ). .. The following primers were used to clone the fragments from genomic tail DNA containing the Mfe I restriction enzyme site and subsequently digested by Mfe I (New England Biolabs).

    other:

    Article Title: Amplified-Fragment Length Polymorphism Fingerprinting of Mycoplasma Species
    Article Snippet: Five microliters of the DNA samples containing approximately 200 to 600 ng of genomic DNA was simultaneously digested with 10 U of Bgl II and 10 U of Mfe I (New England Biolabs, Beverly, Mass.) at 37°C for 2 h in a restriction buffer containing 10 mM Tris-acetate (pH 7.5), 10 mM Mg acetate, 50 mM K acetate, 5 mM dithiothreitol, and 50 ng of bovine serum albumin per μl ( ).

    Article Title: Intestinal carriage of Campylobacter jejuni and Campylobacter coli among cattle from South-western Norway and comparative genotyping of bovine and human isolates by amplified-fragment length polymorphism
    Article Snippet: The AFLP analysis was performed as detailed by Lindstedt et al. (2000), using the restriction enzymes Bgl II and Mfe I (New England Biolabs, Beverly, MA, USA).

    Polymerase Chain Reaction:

    Article Title: Small- and Large-Effect Quantitative Trait Locus Interactions Underlie Variation in Yeast Sporulation Efficiency
    Article Snippet: In 96 well plates, approximately 0.3 μg of DNA from each strain was digested with 5 units each of Mfe I and Mbo I [New England Biolabs (NEB)] in NEB buffer 4 for 1 hr at 37°, then heat inactivated for 20 min at 65°. .. Fifteen microliters of each ligated sample was pooled into groups of 48 containing 44 offspring and 2 of each parent as controls and purified using the QIAquick PCR purification kit (Qiagen).

    Article Title: Haemophilus parainfluenzae expresses diverse lipopolysaccharide O-antigens using ABC transporter and Wzy polymerase-dependent mechanisms
    Article Snippet: Paragraph title: Long range PCR (LR-PCR) and digests ... Five microlitres of each LR-PCR product was digested with MfeI (NEB).

    Article Title: Hygromycin B and Apramycin Antibiotic Resistance Cassettes for Use in Campylobacter jejuni
    Article Snippet: .. The initial aph (7″) PCR product was instead digested with Mfe I and Sph I (NEB), purified, and ligated to low-copy pBAD24 digested with Eco RI and Sph I. .. The ligation was transformed into E. coli DH5α and sequencing of the transformants indicated the correct aph (7″) sequence was incorporated.

    Article Title: A Novel Mechanism of Resistance to Mouse Mammary Tumor Virus Infection
    Article Snippet: Southern blots of PCR products were hybridized with LTR-specific probe as previously published ( I (previously described (24). .. After amplification and agarose gel electrophoresis, the products were transferred to nylon membranes and hybridized with a probe specific to the MMTV LTR ( Semiquantitative RT-PCR analysis was accomplished by synthesis of cDNA from RNA isolated from viral particles with MMTV(C3H) LTR-specific primers, followed by digestion with Mfe I (New England Biolabs, Beverly, Mass.) as previously described ( ).

    Pulsed-Field Gel:

    Article Title: Restriction Endonucleases from Invasive Neisseria gonorrhoeae Cause Double-Strand Breaks and Distort Mitosis in Epithelial Cells during Infection
    Article Snippet: Paragraph title: Pulsed-field gel electrophoresis ... As positive controls, 0.5 µl each of the restriction enzymes NgoMIV, BamHI, KpnI and MfeI (New England Biolabs, Ipswich, MA, USA) and 5 µl CutSmart buffer were added to 1 µg genomic DNA in 50 µl reactions.

    BAC Assay:

    Article Title: Enhanced Striatal Dopamine Transmission and Motor Performance with LRRK2 Overexpression in Mice is Eliminated by familial Parkinson's Disease Mutation G2019S
    Article Snippet: Paragraph title: Generation of LRRK2-Wt and LRRK2-G2019S BAC transgenic mice ... The following primers were used to clone the fragments from genomic tail DNA containing the Mfe I restriction enzyme site and subsequently digested by Mfe I (New England Biolabs).

    Mutagenesis:

    Article Title: Enhanced Striatal Dopamine Transmission and Motor Performance with LRRK2 Overexpression in Mice is Eliminated by familial Parkinson's Disease Mutation G2019S
    Article Snippet: As a result of this sequence change, Mfe I restriction enzyme site was created for the confirmation of mutation in genotyping. .. The following primers were used to clone the fragments from genomic tail DNA containing the Mfe I restriction enzyme site and subsequently digested by Mfe I (New England Biolabs).

    Isolation:

    Article Title: A Novel Mechanism of Resistance to Mouse Mammary Tumor Virus Infection
    Article Snippet: .. After amplification and agarose gel electrophoresis, the products were transferred to nylon membranes and hybridized with a probe specific to the MMTV LTR ( Semiquantitative RT-PCR analysis was accomplished by synthesis of cDNA from RNA isolated from viral particles with MMTV(C3H) LTR-specific primers, followed by digestion with Mfe I (New England Biolabs, Beverly, Mass.) as previously described ( ). .. After amplification and agarose gel electrophoresis, the products were transferred to nylon membranes and hybridized with a probe specific to the MMTV LTR ( ).

    Article Title: The Transcriptional Enhancer of the Pea Plastocyanin Gene Associates with the Nuclear Matrix and Regulates Gene Expression through Histone Acetylation
    Article Snippet: Nuclei were isolated from 10 g of tobacco leaf tissues as described previously and suspended in 10 mL of NSB. .. After six washes with buffer D (20 mM Tris-HCl, pH 8.0, 70 mM NaCl, 20 mM KCl, 10 mM MgCl2 , 0.125 mM spermidine, 0.05 mM spermine, and 0.1% digitonin), the matrices were suspended in 500 μL of buffer D, and 200 units of PvuII, MfeI, and SacI (New England Biolabs, Beverly, MA) were added.

    Article Title: Enhanced Striatal Dopamine Transmission and Motor Performance with LRRK2 Overexpression in Mice is Eliminated by familial Parkinson's Disease Mutation G2019S
    Article Snippet: The following primers were used to clone the fragments from genomic tail DNA containing the Mfe I restriction enzyme site and subsequently digested by Mfe I (New England Biolabs). .. The following primers were used to amplify the kinase domain by RT-PCR using mRNA isolated from transgenic and control brains and the cDNA was purified and digested by Mfe I .

    Mouse Assay:

    Article Title: A Novel Mechanism of Resistance to Mouse Mammary Tumor Virus Infection
    Article Snippet: Primary lymphocytes isolated from spleens of two (see Fig. B) or four (Fig. B) 4-month-old MMTV-infected I/LnJ and C3H/HeN mice were plated at 4 · 106 cells/ml in Click's medium (Life Technologies) supplemented with 5% fetal calf serum in the presence of 1 μg of lipopolysaccharide (Sigma, Inc., St. Louis, Mo.) per ml. .. After amplification and agarose gel electrophoresis, the products were transferred to nylon membranes and hybridized with a probe specific to the MMTV LTR ( Semiquantitative RT-PCR analysis was accomplished by synthesis of cDNA from RNA isolated from viral particles with MMTV(C3H) LTR-specific primers, followed by digestion with Mfe I (New England Biolabs, Beverly, Mass.) as previously described ( ).

    Article Title: Enhanced Striatal Dopamine Transmission and Motor Performance with LRRK2 Overexpression in Mice is Eliminated by familial Parkinson's Disease Mutation G2019S
    Article Snippet: Paragraph title: Generation of LRRK2-Wt and LRRK2-G2019S BAC transgenic mice ... The following primers were used to clone the fragments from genomic tail DNA containing the Mfe I restriction enzyme site and subsequently digested by Mfe I (New England Biolabs).

    Sequencing:

    Article Title: Small- and Large-Effect Quantitative Trait Locus Interactions Underlie Variation in Yeast Sporulation Efficiency
    Article Snippet: In 96 well plates, approximately 0.3 μg of DNA from each strain was digested with 5 units each of Mfe I and Mbo I [New England Biolabs (NEB)] in NEB buffer 4 for 1 hr at 37°, then heat inactivated for 20 min at 65°. .. The digested DNA from each strain to be genotyped was ligated to one pair of modified Illumina sequencing adapters (IDT, sequences available in supporting information, ) containing 1 of 48 different 4-bp barcodes using 1000 units of T4 ligase (NEB) in 1× ligation buffer containing dATP.

    Article Title: CRISPR-Cas9 fusion to dominant-negative 53BP1 enhances HDR and inhibits NHEJ specifically at Cas9 target sites
    Article Snippet: For the tNGFR Cas9-DN1S construct, SpCas9.TGS.DN1S was moved from the PX458M plasmid construct into the CMV.tNGFR lentiviral construct between NcoI and MfeI (NEB). .. Clones were screened for the correct sequence, then used to generate dSpCas9 and dSpCas9-DN1S constructs without gRNA and with gRNA to target the human CD45 locus.

    Article Title: Hygromycin B and Apramycin Antibiotic Resistance Cassettes for Use in Campylobacter jejuni
    Article Snippet: Sequencing of pGEM-T containing aph (7″) indicated that the restriction sites flanking aph (7″) and aph (7″) sequence itself were incorrect. .. The initial aph (7″) PCR product was instead digested with Mfe I and Sph I (NEB), purified, and ligated to low-copy pBAD24 digested with Eco RI and Sph I.

    Article Title: Enhanced Striatal Dopamine Transmission and Motor Performance with LRRK2 Overexpression in Mice is Eliminated by familial Parkinson's Disease Mutation G2019S
    Article Snippet: As a result of this sequence change, Mfe I restriction enzyme site was created for the confirmation of mutation in genotyping. .. The following primers were used to clone the fragments from genomic tail DNA containing the Mfe I restriction enzyme site and subsequently digested by Mfe I (New England Biolabs).

    Purification:

    Article Title: Small- and Large-Effect Quantitative Trait Locus Interactions Underlie Variation in Yeast Sporulation Efficiency
    Article Snippet: In 96 well plates, approximately 0.3 μg of DNA from each strain was digested with 5 units each of Mfe I and Mbo I [New England Biolabs (NEB)] in NEB buffer 4 for 1 hr at 37°, then heat inactivated for 20 min at 65°. .. Fifteen microliters of each ligated sample was pooled into groups of 48 containing 44 offspring and 2 of each parent as controls and purified using the QIAquick PCR purification kit (Qiagen).

    Article Title: Hygromycin B and Apramycin Antibiotic Resistance Cassettes for Use in Campylobacter jejuni
    Article Snippet: .. The initial aph (7″) PCR product was instead digested with Mfe I and Sph I (NEB), purified, and ligated to low-copy pBAD24 digested with Eco RI and Sph I. .. The ligation was transformed into E. coli DH5α and sequencing of the transformants indicated the correct aph (7″) sequence was incorporated.

    Article Title: A Novel Mechanism of Resistance to Mouse Mammary Tumor Virus Infection
    Article Snippet: Paragraph title: Virus purification and reverse transcription (RT)-PCR analysis. ... After amplification and agarose gel electrophoresis, the products were transferred to nylon membranes and hybridized with a probe specific to the MMTV LTR ( Semiquantitative RT-PCR analysis was accomplished by synthesis of cDNA from RNA isolated from viral particles with MMTV(C3H) LTR-specific primers, followed by digestion with Mfe I (New England Biolabs, Beverly, Mass.) as previously described ( ).

    Article Title: Enhanced Striatal Dopamine Transmission and Motor Performance with LRRK2 Overexpression in Mice is Eliminated by familial Parkinson's Disease Mutation G2019S
    Article Snippet: The following primers were used to clone the fragments from genomic tail DNA containing the Mfe I restriction enzyme site and subsequently digested by Mfe I (New England Biolabs). .. The following primers were used to amplify the kinase domain by RT-PCR using mRNA isolated from transgenic and control brains and the cDNA was purified and digested by Mfe I .

    Plasmid Preparation:

    Article Title: CRISPR-Cas9 fusion to dominant-negative 53BP1 enhances HDR and inhibits NHEJ specifically at Cas9 target sites
    Article Snippet: .. For the tNGFR Cas9-DN1S construct, SpCas9.TGS.DN1S was moved from the PX458M plasmid construct into the CMV.tNGFR lentiviral construct between NcoI and MfeI (NEB). .. Both DN1S and Cas9-DN1S tNGFR constructs were driven by the MND promoter.

    Article Title: Hygromycin B and Apramycin Antibiotic Resistance Cassettes for Use in Campylobacter jejuni
    Article Snippet: Sequencing verified that the fragment containing aac (3)IV was correctly inserted into pGEM-T and this plasmid was then designated pAC1A. .. The initial aph (7″) PCR product was instead digested with Mfe I and Sph I (NEB), purified, and ligated to low-copy pBAD24 digested with Eco RI and Sph I.

    Article Title: The β3‐integrin endothelial adhesome regulates microtubule‐dependent cell migration
    Article Snippet: Generation of human β3 integrin expressing cells 1 × 106 β3NULL endothelial cells were transfected with 10 μg of MfeI (New England Biolabs, Hitchin, UK) linearised full‐length human β3‐integrin (see Robinson et al ) cloned into pcDNA™ 6.2/C‐EmGFP (see Amaxa nucleofection above). .. An empty vector (EV) was used as a control.

    Agarose Gel Electrophoresis:

    Article Title: A Novel Mechanism of Resistance to Mouse Mammary Tumor Virus Infection
    Article Snippet: .. After amplification and agarose gel electrophoresis, the products were transferred to nylon membranes and hybridized with a probe specific to the MMTV LTR ( Semiquantitative RT-PCR analysis was accomplished by synthesis of cDNA from RNA isolated from viral particles with MMTV(C3H) LTR-specific primers, followed by digestion with Mfe I (New England Biolabs, Beverly, Mass.) as previously described ( ). .. After amplification and agarose gel electrophoresis, the products were transferred to nylon membranes and hybridized with a probe specific to the MMTV LTR ( ).

    Article Title: The Transcriptional Enhancer of the Pea Plastocyanin Gene Associates with the Nuclear Matrix and Regulates Gene Expression through Histone Acetylation
    Article Snippet: The DNA was separated on a 1% agarose gel, which was fixed in 7% trichloroacetic acid for 30 min, dried, and autoradiographed. .. After six washes with buffer D (20 mM Tris-HCl, pH 8.0, 70 mM NaCl, 20 mM KCl, 10 mM MgCl2 , 0.125 mM spermidine, 0.05 mM spermine, and 0.1% digitonin), the matrices were suspended in 500 μL of buffer D, and 200 units of PvuII, MfeI, and SacI (New England Biolabs, Beverly, MA) were added.

    Transgenic Assay:

    Article Title: Enhanced Striatal Dopamine Transmission and Motor Performance with LRRK2 Overexpression in Mice is Eliminated by familial Parkinson's Disease Mutation G2019S
    Article Snippet: Paragraph title: Generation of LRRK2-Wt and LRRK2-G2019S BAC transgenic mice ... The following primers were used to clone the fragments from genomic tail DNA containing the Mfe I restriction enzyme site and subsequently digested by Mfe I (New England Biolabs).

    Ethanol Precipitation:

    Article Title: The Transcriptional Enhancer of the Pea Plastocyanin Gene Associates with the Nuclear Matrix and Regulates Gene Expression through Histone Acetylation
    Article Snippet: Nuclear matrices were collected by centrifugation, and both the supernatant and pellet fractions were treated with proteinase K at 1 mg/mL in 1% SDS at 65°C for 1 h. DNA was extracted from the pellet and supernatant fractions by phenol/chloroform extraction and ethanol precipitation. .. After six washes with buffer D (20 mM Tris-HCl, pH 8.0, 70 mM NaCl, 20 mM KCl, 10 mM MgCl2 , 0.125 mM spermidine, 0.05 mM spermine, and 0.1% digitonin), the matrices were suspended in 500 μL of buffer D, and 200 units of PvuII, MfeI, and SacI (New England Biolabs, Beverly, MA) were added.

    Produced:

    Article Title: CRISPR-Cas9 fusion to dominant-negative 53BP1 enhances HDR and inhibits NHEJ specifically at Cas9 target sites
    Article Snippet: Lentivirus was produced as described above. .. For the tNGFR Cas9-DN1S construct, SpCas9.TGS.DN1S was moved from the PX458M plasmid construct into the CMV.tNGFR lentiviral construct between NcoI and MfeI (NEB).

    Article Title: A Simple Genotyping Method for Rapid Differentiation of Blastocystis Subtypes and Subtype Distribution of Blastocystis spp. in Thailand
    Article Snippet: In step 1 digestion, the 480 bp-PCR amplicons were digested separately with Mfe I and Ase I enzyme (New England BioLabs, Pickering, ON, Canada). .. The digestion produced 5 RFLP patterns (Pattern A–E) ( ).

    Concentration Assay:

    Article Title: Small- and Large-Effect Quantitative Trait Locus Interactions Underlie Variation in Yeast Sporulation Efficiency
    Article Snippet: In 96 well plates, approximately 0.3 μg of DNA from each strain was digested with 5 units each of Mfe I and Mbo I [New England Biolabs (NEB)] in NEB buffer 4 for 1 hr at 37°, then heat inactivated for 20 min at 65°. .. Adapters were preannealed and added so that the final concentration was 100 nM for sequencing adapter (P1) and 10 µM for secondary adapter (P2).

    DNA Purification:

    Article Title: Restriction Endonucleases from Invasive Neisseria gonorrhoeae Cause Double-Strand Breaks and Distort Mitosis in Epithelial Cells during Infection
    Article Snippet: Pulsed-field gel electrophoresis Genomic DNA from VK2/E6E7 cells was extracted using the Wizard Genomic DNA Purification Kit (Promega, Madison, WI, USA), according to the manufacturer's recommendations. .. As positive controls, 0.5 µl each of the restriction enzymes NgoMIV, BamHI, KpnI and MfeI (New England Biolabs, Ipswich, MA, USA) and 5 µl CutSmart buffer were added to 1 µg genomic DNA in 50 µl reactions.

    Marker:

    Article Title: Hygromycin B and Apramycin Antibiotic Resistance Cassettes for Use in Campylobacter jejuni
    Article Snippet: The initial aph (7″) PCR product was instead digested with Mfe I and Sph I (NEB), purified, and ligated to low-copy pBAD24 digested with Eco RI and Sph I. .. The resulting plasmid with the aph (7″) non-polar marker inserted in pBAD24 was designated pAC1H.

    Staining:

    Article Title: Restriction Endonucleases from Invasive Neisseria gonorrhoeae Cause Double-Strand Breaks and Distort Mitosis in Epithelial Cells during Infection
    Article Snippet: As positive controls, 0.5 µl each of the restriction enzymes NgoMIV, BamHI, KpnI and MfeI (New England Biolabs, Ipswich, MA, USA) and 5 µl CutSmart buffer were added to 1 µg genomic DNA in 50 µl reactions. .. DNA was separated (120° field angle, 240 s switch time, 4 V/cm, 14°C) for 20 h. Gels was stained with ethidium bromide overnight at 4°C and subsequently examined in a UV illuminator.

    Gradient Centrifugation:

    Article Title: A Novel Mechanism of Resistance to Mouse Mammary Tumor Virus Infection
    Article Snippet: RNA was extracted from the virus pellet by guanidine thiocyanate extraction and CsCl gradient centrifugation ( ). .. After amplification and agarose gel electrophoresis, the products were transferred to nylon membranes and hybridized with a probe specific to the MMTV LTR ( Semiquantitative RT-PCR analysis was accomplished by synthesis of cDNA from RNA isolated from viral particles with MMTV(C3H) LTR-specific primers, followed by digestion with Mfe I (New England Biolabs, Beverly, Mass.) as previously described ( ).

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  • 90
    New England Biolabs mfei restriction enzyme
    Mfei Restriction Enzyme, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mfei restriction enzyme/product/New England Biolabs
    Average 90 stars, based on 24 article reviews
    Price from $9.99 to $1999.99
    mfei restriction enzyme - by Bioz Stars, 2020-04
    90/100 stars
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