gl 2 deletion virus 79vb4 cells  (New England Biolabs)


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    Name:
    BmgBI
    Description:
    BmgBI 2 500 units
    Catalog Number:
    r0628l
    Price:
    290
    Size:
    2 500 units
    Category:
    Restriction Enzymes
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    Structured Review

    New England Biolabs gl 2 deletion virus 79vb4 cells
    BmgBI
    BmgBI 2 500 units
    https://www.bioz.com/result/gl 2 deletion virus 79vb4 cells/product/New England Biolabs
    Average 92 stars, based on 13 article reviews
    Price from $9.99 to $1999.99
    gl 2 deletion virus 79vb4 cells - by Bioz Stars, 2020-09
    92/100 stars

    Images

    1) Product Images from "Herpes simplex viruses activate phospholipid scramblase to redistribute phosphatidylserines and Akt to the outer leaflet of the plasma membrane and promote viral entry"

    Article Title: Herpes simplex viruses activate phospholipid scramblase to redistribute phosphatidylserines and Akt to the outer leaflet of the plasma membrane and promote viral entry

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1006766

    Phospholipid scramblase associates with glycoprotein L to restore membrane architecture. (A) CaSki cells were mock-infected or synchronously infected with HSV-2(G), HSV-1(KOS), complemented or non-complemented gL-2 deletion virus (ΔgL-2 +/- and ΔgL-2 -/- , respectively), non-complemented gH-2 deletion (ΔgH-2 -/- ), or non-complemented gL-1 (ΔgL-1 -/- ) or gH-1 deleted viruses (ΔgH-1 -/- ) (MOI equivalent to ~ 1 pfu/cell). Fifteen minutes after the temperature shift, the cells were fixed and stained with the indicated murine HSV-serotype common mAbs and a rabbit polyclonal antibody against PLSCR1 and then probed with species-specific proximity ligation secondary antibodies. Results are representative of at least 3 independent experiments. (B). Western blots of dextran gradient-purified ΔgL or ΔgH virus isolated 24 hours after infection of 79VB4 (ΔgL-2 +/- ), F6 (ΔgH-2 +/- ) or Vero (ΔgL-2 −/− and ΔgH-2 -/- ) cells. Protein expression was assessed for viral glycoproteins H and L with serotype-specific mAbs. (C). CaSki cells were synchronously infected with HSV-2(G) (5 pfu/cell) (+) or mock-infected (-) and cell lysates were prepared 15 min post-temperature shift, immunoprecipitated (IP) with serotype common mouse anti-gL, mouse ant-gH or rabbit anti-PLSCR1 and equivalent volumes of supernatant, pellet or whole cell lysates analyzed by preparing Western blots (WB) and probing with rabbit anti-PLSCR1, mouse anti-gL, mouse anti-gH or mouse anti-αvβ3. Blots are representative of results obtained in 2 independent experiments. (D). CaSki cells were infected with HSV-2(G) (MOI 10 pfu/cell) and at the indicated times post-infection, cells were fixed and stained with antibodies to phosphatidylserine (red) or Akt (green); nuclei were stained blue. Mock-infected cells were included as a negative control. Representative extended focus images from two experiments are shown; scale bar 10μm. (E). CaSki cells were infected with HSV-2(G) (MOI 10 pfu/cell) for 30 minutes and then the inoculum removed, cells washed with a low pH citrate buffer and then incubated for 1, 2 or 4 h. R5421 (or DMSO) was added to the medium at the time of infection (t = 0 minutes) or immediately following citrate treatment (t = 30 minutes). Cells were stained as in Panel D. Representative images from two experiments are shown; bar = 10μm.
    Figure Legend Snippet: Phospholipid scramblase associates with glycoprotein L to restore membrane architecture. (A) CaSki cells were mock-infected or synchronously infected with HSV-2(G), HSV-1(KOS), complemented or non-complemented gL-2 deletion virus (ΔgL-2 +/- and ΔgL-2 -/- , respectively), non-complemented gH-2 deletion (ΔgH-2 -/- ), or non-complemented gL-1 (ΔgL-1 -/- ) or gH-1 deleted viruses (ΔgH-1 -/- ) (MOI equivalent to ~ 1 pfu/cell). Fifteen minutes after the temperature shift, the cells were fixed and stained with the indicated murine HSV-serotype common mAbs and a rabbit polyclonal antibody against PLSCR1 and then probed with species-specific proximity ligation secondary antibodies. Results are representative of at least 3 independent experiments. (B). Western blots of dextran gradient-purified ΔgL or ΔgH virus isolated 24 hours after infection of 79VB4 (ΔgL-2 +/- ), F6 (ΔgH-2 +/- ) or Vero (ΔgL-2 −/− and ΔgH-2 -/- ) cells. Protein expression was assessed for viral glycoproteins H and L with serotype-specific mAbs. (C). CaSki cells were synchronously infected with HSV-2(G) (5 pfu/cell) (+) or mock-infected (-) and cell lysates were prepared 15 min post-temperature shift, immunoprecipitated (IP) with serotype common mouse anti-gL, mouse ant-gH or rabbit anti-PLSCR1 and equivalent volumes of supernatant, pellet or whole cell lysates analyzed by preparing Western blots (WB) and probing with rabbit anti-PLSCR1, mouse anti-gL, mouse anti-gH or mouse anti-αvβ3. Blots are representative of results obtained in 2 independent experiments. (D). CaSki cells were infected with HSV-2(G) (MOI 10 pfu/cell) and at the indicated times post-infection, cells were fixed and stained with antibodies to phosphatidylserine (red) or Akt (green); nuclei were stained blue. Mock-infected cells were included as a negative control. Representative extended focus images from two experiments are shown; scale bar 10μm. (E). CaSki cells were infected with HSV-2(G) (MOI 10 pfu/cell) for 30 minutes and then the inoculum removed, cells washed with a low pH citrate buffer and then incubated for 1, 2 or 4 h. R5421 (or DMSO) was added to the medium at the time of infection (t = 0 minutes) or immediately following citrate treatment (t = 30 minutes). Cells were stained as in Panel D. Representative images from two experiments are shown; bar = 10μm.

    Techniques Used: Infection, Staining, Ligation, Western Blot, Purification, Isolation, Expressing, Immunoprecipitation, Negative Control, Incubation

    2) Product Images from "Herpes simplex viruses activate phospholipid scramblase to redistribute phosphatidylserines and Akt to the outer leaflet of the plasma membrane and promote viral entry"

    Article Title: Herpes simplex viruses activate phospholipid scramblase to redistribute phosphatidylserines and Akt to the outer leaflet of the plasma membrane and promote viral entry

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1006766

    Phospholipid scramblase associates with glycoprotein L to restore membrane architecture. (A) CaSki cells were mock-infected or synchronously infected with HSV-2(G), HSV-1(KOS), complemented or non-complemented gL-2 deletion virus (ΔgL-2 +/- and ΔgL-2 -/- , respectively), non-complemented gH-2 deletion (ΔgH-2 -/- ), or non-complemented gL-1 (ΔgL-1 -/- ) or gH-1 deleted viruses (ΔgH-1 -/- ) (MOI equivalent to ~ 1 pfu/cell). Fifteen minutes after the temperature shift, the cells were fixed and stained with the indicated murine HSV-serotype common mAbs and a rabbit polyclonal antibody against PLSCR1 and then probed with species-specific proximity ligation secondary antibodies. Results are representative of at least 3 independent experiments. (B). Western blots of dextran gradient-purified ΔgL or ΔgH virus isolated 24 hours after infection of 79VB4 (ΔgL-2 +/- ), F6 (ΔgH-2 +/- ) or Vero (ΔgL-2 −/− and ΔgH-2 -/- ) cells. Protein expression was assessed for viral glycoproteins H and L with serotype-specific mAbs. (C). CaSki cells were synchronously infected with HSV-2(G) (5 pfu/cell) (+) or mock-infected (-) and cell lysates were prepared 15 min post-temperature shift, immunoprecipitated (IP) with serotype common mouse anti-gL, mouse ant-gH or rabbit anti-PLSCR1 and equivalent volumes of supernatant, pellet or whole cell lysates analyzed by preparing Western blots (WB) and probing with rabbit anti-PLSCR1, mouse anti-gL, mouse anti-gH or mouse anti-αvβ3. Blots are representative of results obtained in 2 independent experiments. (D). CaSki cells were infected with HSV-2(G) (MOI 10 pfu/cell) and at the indicated times post-infection, cells were fixed and stained with antibodies to phosphatidylserine (red) or Akt (green); nuclei were stained blue. Mock-infected cells were included as a negative control. Representative extended focus images from two experiments are shown; scale bar 10μm. (E). CaSki cells were infected with HSV-2(G) (MOI 10 pfu/cell) for 30 minutes and then the inoculum removed, cells washed with a low pH citrate buffer and then incubated for 1, 2 or 4 h. R5421 (or DMSO) was added to the medium at the time of infection (t = 0 minutes) or immediately following citrate treatment (t = 30 minutes). Cells were stained as in Panel D. Representative images from two experiments are shown; bar = 10μm.
    Figure Legend Snippet: Phospholipid scramblase associates with glycoprotein L to restore membrane architecture. (A) CaSki cells were mock-infected or synchronously infected with HSV-2(G), HSV-1(KOS), complemented or non-complemented gL-2 deletion virus (ΔgL-2 +/- and ΔgL-2 -/- , respectively), non-complemented gH-2 deletion (ΔgH-2 -/- ), or non-complemented gL-1 (ΔgL-1 -/- ) or gH-1 deleted viruses (ΔgH-1 -/- ) (MOI equivalent to ~ 1 pfu/cell). Fifteen minutes after the temperature shift, the cells were fixed and stained with the indicated murine HSV-serotype common mAbs and a rabbit polyclonal antibody against PLSCR1 and then probed with species-specific proximity ligation secondary antibodies. Results are representative of at least 3 independent experiments. (B). Western blots of dextran gradient-purified ΔgL or ΔgH virus isolated 24 hours after infection of 79VB4 (ΔgL-2 +/- ), F6 (ΔgH-2 +/- ) or Vero (ΔgL-2 −/− and ΔgH-2 -/- ) cells. Protein expression was assessed for viral glycoproteins H and L with serotype-specific mAbs. (C). CaSki cells were synchronously infected with HSV-2(G) (5 pfu/cell) (+) or mock-infected (-) and cell lysates were prepared 15 min post-temperature shift, immunoprecipitated (IP) with serotype common mouse anti-gL, mouse ant-gH or rabbit anti-PLSCR1 and equivalent volumes of supernatant, pellet or whole cell lysates analyzed by preparing Western blots (WB) and probing with rabbit anti-PLSCR1, mouse anti-gL, mouse anti-gH or mouse anti-αvβ3. Blots are representative of results obtained in 2 independent experiments. (D). CaSki cells were infected with HSV-2(G) (MOI 10 pfu/cell) and at the indicated times post-infection, cells were fixed and stained with antibodies to phosphatidylserine (red) or Akt (green); nuclei were stained blue. Mock-infected cells were included as a negative control. Representative extended focus images from two experiments are shown; scale bar 10μm. (E). CaSki cells were infected with HSV-2(G) (MOI 10 pfu/cell) for 30 minutes and then the inoculum removed, cells washed with a low pH citrate buffer and then incubated for 1, 2 or 4 h. R5421 (or DMSO) was added to the medium at the time of infection (t = 0 minutes) or immediately following citrate treatment (t = 30 minutes). Cells were stained as in Panel D. Representative images from two experiments are shown; bar = 10μm.

    Techniques Used: Infection, Staining, Ligation, Western Blot, Purification, Isolation, Expressing, Immunoprecipitation, Negative Control, Incubation

    3) Product Images from "Genetic Characterization of Second-Line Drug-Resistant and Extensively Drug-Resistant Mycobacterium tuberculosis from the Northern Region of India"

    Article Title: Genetic Characterization of Second-Line Drug-Resistant and Extensively Drug-Resistant Mycobacterium tuberculosis from the Northern Region of India

    Journal: Journal of Epidemiology and Global Health

    doi: 10.2991/j.jegh.2018.02.100

    PCR RFLP of rrs gene after digestion with BmgB I. Lane 1, 100 bp DNA ladder; Lanes 2 and 3, 353 and 128 bp fragments with wild-type sequence; and Lanes 4 and 5, 481 bp fragments of mutant sequence
    Figure Legend Snippet: PCR RFLP of rrs gene after digestion with BmgB I. Lane 1, 100 bp DNA ladder; Lanes 2 and 3, 353 and 128 bp fragments with wild-type sequence; and Lanes 4 and 5, 481 bp fragments of mutant sequence

    Techniques Used: Polymerase Chain Reaction, Sequencing, Mutagenesis

    Related Articles

    Combined Bisulfite Restriction Analysis Assay:

    Article Title: Evidence for high bi-allelic expression of activating Ly49 receptors
    Article Snippet: .. Combined bisulfite and restriction enzyme analysis For COBRA, gel purified fragments of Ly49d upstream and downstream regions were digested with Taqα I and BmgBI restriction enzyme (NEB), respectively to distinguish between methylated (CpG) and unmethylated (TpG). .. Only fragments that were originally methylated in the gDNA and therefore not converted by sodium bisulfite treatment are cut.

    Amplification:

    Article Title: Novel Acidic Sesquiterpenoids Constitute a Dominant Class of Pathogen-Induced Phytoalexins in Maize 1Novel Acidic Sesquiterpenoids Constitute a Dominant Class of Pathogen-Induced Phytoalexins in Maize 1 [W]Novel Acidic Sesquiterpenoids Constitute a Dominant Class of Pathogen-Induced Phytoalexins in Maize 1 [W] [OA]
    Article Snippet: .. To confirm the specificity of Tps6 and Tps11 primers, the PCR products of each primer pair were digested with BmgBI (New England Biolabs), which specifically digests the Tps6 amplicon but not Tps11. ..

    Ligation:

    Article Title: Functional Analysis of a Novel cis-Acting Regulatory Region within the Human Ankyrin Gene (ANK-1) Promoter ▿
    Article Snippet: .. The PCR products were digested with BmgB1 and XhoI and ligated into the pGL2B-based ANK-1 /luciferase plasmid described previously (pANK-1WT ; p296 from reference ) by using a quick ligation kit (New England Biolabs, Ipswich, MA). .. Maximum-efficiency DH5α bacteria (Invitrogen, Gaithersburg, MD) were transformed with the ligation products and grown for 16 h in LB medium containing ampicillin, after which plasmid DNA was prepared.

    Selection:

    Article Title: Herpes simplex viruses activate phospholipid scramblase to redistribute phosphatidylserines and Akt to the outer leaflet of the plasma membrane and promote viral entry
    Article Snippet: .. Construction of a gL-2 deletion virus 79VB4 cells were co-transfected with HSV-2(G) DNA and DNA from an engineered plasmid (designated pEF6/5UL1HygroGFP) that contains the gL-2 gene (UL1) disrupted at the BmgBI and HpaI sites (R0628, R0105, New England BioLabs) by the hygromycin enhanced green fluorescent protein (EGFP) fusion protein under the control of the immediate early promoter of human cytomegalovirus (HCMV) constructed from pHygGFP (Clontech) (The plasmid encoding HSV-2(G) gL-2, designated pEF6/5 UL1 was a gift from Torin Weisbrod and William Jacobs).This dual function marker vector allows for drug selection with the ability to identify positive transfectants using GFP as a fluorescent reporter. .. Cotransfections were performed using the Effectene Transfection reagent (301425, Qiagen).

    Article Title: Herpes simplex viruses activate phospholipid scramblase to redistribute phosphatidylserines and Akt to the outer leaflet of the plasma membrane and promote viral entry
    Article Snippet: .. 79VB4 cells were co-transfected with HSV-2(G) DNA and DNA from an engineered plasmid (designated pEF6/5UL1HygroGFP) that contains the gL-2 gene (UL1) disrupted at the BmgBI and HpaI sites (R0628, R0105, New England BioLabs) by the hygromycin enhanced green fluorescent protein (EGFP) fusion protein under the control of the immediate early promoter of human cytomegalovirus (HCMV) constructed from pHygGFP (Clontech) (The plasmid encoding HSV-2(G) gL-2, designated pEF6/5 UL1 was a gift from Torin Weisbrod and William Jacobs).This dual function marker vector allows for drug selection with the ability to identify positive transfectants using GFP as a fluorescent reporter. .. Cotran sfections were performed using the Effectene Transfection reagent (301425, Qiagen).

    Methylation:

    Article Title: Evidence for high bi-allelic expression of activating Ly49 receptors
    Article Snippet: .. Combined bisulfite and restriction enzyme analysis For COBRA, gel purified fragments of Ly49d upstream and downstream regions were digested with Taqα I and BmgBI restriction enzyme (NEB), respectively to distinguish between methylated (CpG) and unmethylated (TpG). .. Only fragments that were originally methylated in the gDNA and therefore not converted by sodium bisulfite treatment are cut.

    Construct:

    Article Title: Herpes simplex viruses activate phospholipid scramblase to redistribute phosphatidylserines and Akt to the outer leaflet of the plasma membrane and promote viral entry
    Article Snippet: .. Construction of a gL-2 deletion virus 79VB4 cells were co-transfected with HSV-2(G) DNA and DNA from an engineered plasmid (designated pEF6/5UL1HygroGFP) that contains the gL-2 gene (UL1) disrupted at the BmgBI and HpaI sites (R0628, R0105, New England BioLabs) by the hygromycin enhanced green fluorescent protein (EGFP) fusion protein under the control of the immediate early promoter of human cytomegalovirus (HCMV) constructed from pHygGFP (Clontech) (The plasmid encoding HSV-2(G) gL-2, designated pEF6/5 UL1 was a gift from Torin Weisbrod and William Jacobs).This dual function marker vector allows for drug selection with the ability to identify positive transfectants using GFP as a fluorescent reporter. .. Cotransfections were performed using the Effectene Transfection reagent (301425, Qiagen).

    Article Title: Herpes simplex viruses activate phospholipid scramblase to redistribute phosphatidylserines and Akt to the outer leaflet of the plasma membrane and promote viral entry
    Article Snippet: .. 79VB4 cells were co-transfected with HSV-2(G) DNA and DNA from an engineered plasmid (designated pEF6/5UL1HygroGFP) that contains the gL-2 gene (UL1) disrupted at the BmgBI and HpaI sites (R0628, R0105, New England BioLabs) by the hygromycin enhanced green fluorescent protein (EGFP) fusion protein under the control of the immediate early promoter of human cytomegalovirus (HCMV) constructed from pHygGFP (Clontech) (The plasmid encoding HSV-2(G) gL-2, designated pEF6/5 UL1 was a gift from Torin Weisbrod and William Jacobs).This dual function marker vector allows for drug selection with the ability to identify positive transfectants using GFP as a fluorescent reporter. .. Cotran sfections were performed using the Effectene Transfection reagent (301425, Qiagen).

    Purification:

    Article Title: Evidence for high bi-allelic expression of activating Ly49 receptors
    Article Snippet: .. Combined bisulfite and restriction enzyme analysis For COBRA, gel purified fragments of Ly49d upstream and downstream regions were digested with Taqα I and BmgBI restriction enzyme (NEB), respectively to distinguish between methylated (CpG) and unmethylated (TpG). .. Only fragments that were originally methylated in the gDNA and therefore not converted by sodium bisulfite treatment are cut.

    Polymerase Chain Reaction:

    Article Title: Novel Acidic Sesquiterpenoids Constitute a Dominant Class of Pathogen-Induced Phytoalexins in Maize 1Novel Acidic Sesquiterpenoids Constitute a Dominant Class of Pathogen-Induced Phytoalexins in Maize 1 [W]Novel Acidic Sesquiterpenoids Constitute a Dominant Class of Pathogen-Induced Phytoalexins in Maize 1 [W] [OA]
    Article Snippet: .. To confirm the specificity of Tps6 and Tps11 primers, the PCR products of each primer pair were digested with BmgBI (New England Biolabs), which specifically digests the Tps6 amplicon but not Tps11. ..

    Article Title: Genetic Characterization of Second-Line Drug-Resistant and Extensively Drug-Resistant Mycobacterium tuberculosis from the Northern Region of India
    Article Snippet: .. For PCR RFLP, the same primers RRSF and RRSR were used followed by the restriction digestion with BmgB I (New England Biolabs, UK). .. Fragments of 353 and 128 bp were obtained from PCR RFLP of rrs gene showing the wild-type genotype ( ).

    Article Title: Functional Analysis of a Novel cis-Acting Regulatory Region within the Human Ankyrin Gene (ANK-1) Promoter ▿
    Article Snippet: .. The PCR products were digested with BmgB1 and XhoI and ligated into the pGL2B-based ANK-1 /luciferase plasmid described previously (pANK-1WT ; p296 from reference ) by using a quick ligation kit (New England Biolabs, Ipswich, MA). .. Maximum-efficiency DH5α bacteria (Invitrogen, Gaithersburg, MD) were transformed with the ligation products and grown for 16 h in LB medium containing ampicillin, after which plasmid DNA was prepared.

    Article Title: Association of A Novel Splice Site Mutation in P/Q-Type Calcium Channels with Childhood Epilepsy and Late-Onset Slowly Progressive Non-Episodic Cerebellar Ataxia
    Article Snippet: .. The resulting PCR product was digested with Dra III and Bmg BI (both from New England Biolabs, Ipswich, MA, USA). .. After gel purification, the fragment was inserted into the corresponding restriction sites of the wild-type CACNA1A construct.

    Luciferase:

    Article Title: Functional Analysis of a Novel cis-Acting Regulatory Region within the Human Ankyrin Gene (ANK-1) Promoter ▿
    Article Snippet: .. The PCR products were digested with BmgB1 and XhoI and ligated into the pGL2B-based ANK-1 /luciferase plasmid described previously (pANK-1WT ; p296 from reference ) by using a quick ligation kit (New England Biolabs, Ipswich, MA). .. Maximum-efficiency DH5α bacteria (Invitrogen, Gaithersburg, MD) were transformed with the ligation products and grown for 16 h in LB medium containing ampicillin, after which plasmid DNA was prepared.

    Marker:

    Article Title: Herpes simplex viruses activate phospholipid scramblase to redistribute phosphatidylserines and Akt to the outer leaflet of the plasma membrane and promote viral entry
    Article Snippet: .. Construction of a gL-2 deletion virus 79VB4 cells were co-transfected with HSV-2(G) DNA and DNA from an engineered plasmid (designated pEF6/5UL1HygroGFP) that contains the gL-2 gene (UL1) disrupted at the BmgBI and HpaI sites (R0628, R0105, New England BioLabs) by the hygromycin enhanced green fluorescent protein (EGFP) fusion protein under the control of the immediate early promoter of human cytomegalovirus (HCMV) constructed from pHygGFP (Clontech) (The plasmid encoding HSV-2(G) gL-2, designated pEF6/5 UL1 was a gift from Torin Weisbrod and William Jacobs).This dual function marker vector allows for drug selection with the ability to identify positive transfectants using GFP as a fluorescent reporter. .. Cotransfections were performed using the Effectene Transfection reagent (301425, Qiagen).

    Article Title: Herpes simplex viruses activate phospholipid scramblase to redistribute phosphatidylserines and Akt to the outer leaflet of the plasma membrane and promote viral entry
    Article Snippet: .. 79VB4 cells were co-transfected with HSV-2(G) DNA and DNA from an engineered plasmid (designated pEF6/5UL1HygroGFP) that contains the gL-2 gene (UL1) disrupted at the BmgBI and HpaI sites (R0628, R0105, New England BioLabs) by the hygromycin enhanced green fluorescent protein (EGFP) fusion protein under the control of the immediate early promoter of human cytomegalovirus (HCMV) constructed from pHygGFP (Clontech) (The plasmid encoding HSV-2(G) gL-2, designated pEF6/5 UL1 was a gift from Torin Weisbrod and William Jacobs).This dual function marker vector allows for drug selection with the ability to identify positive transfectants using GFP as a fluorescent reporter. .. Cotran sfections were performed using the Effectene Transfection reagent (301425, Qiagen).

    other:

    Article Title: Micromechanics of human mitotic chromosomes
    Article Snippet: Restriction enzymes (AluI, RsaI, HaeIII, SnaBI, PvuII, BmgBI, New England Biolabs) were diluted to 0.5 units/μl for microspray experiments, in the buffers recommended by New England Biolabs (NEB): PvuII uses NEB Buffer 2 (50 mM NaCl, 10 mM Tris-HCl, 10mM MgCl2 , 1 mM DTT, pH 7.9); BmgBI uses NEB Buffer 3 (100 mM NaCl, 50 mM Tris-HCl, 10 mM MgCl2 , 1 mM DTT, pH 7.9); AluI, RsaI, HaeIII, SnaBI use NEB Buffer 4 (20 mM Tris-acetate, 50 mM potassium acetate, 10 mM Mg acetate, 1 mM DTT, pH 7.9).

    Plasmid Preparation:

    Article Title: Herpes simplex viruses activate phospholipid scramblase to redistribute phosphatidylserines and Akt to the outer leaflet of the plasma membrane and promote viral entry
    Article Snippet: .. Construction of a gL-2 deletion virus 79VB4 cells were co-transfected with HSV-2(G) DNA and DNA from an engineered plasmid (designated pEF6/5UL1HygroGFP) that contains the gL-2 gene (UL1) disrupted at the BmgBI and HpaI sites (R0628, R0105, New England BioLabs) by the hygromycin enhanced green fluorescent protein (EGFP) fusion protein under the control of the immediate early promoter of human cytomegalovirus (HCMV) constructed from pHygGFP (Clontech) (The plasmid encoding HSV-2(G) gL-2, designated pEF6/5 UL1 was a gift from Torin Weisbrod and William Jacobs).This dual function marker vector allows for drug selection with the ability to identify positive transfectants using GFP as a fluorescent reporter. .. Cotransfections were performed using the Effectene Transfection reagent (301425, Qiagen).

    Article Title: Functional Analysis of a Novel cis-Acting Regulatory Region within the Human Ankyrin Gene (ANK-1) Promoter ▿
    Article Snippet: .. The PCR products were digested with BmgB1 and XhoI and ligated into the pGL2B-based ANK-1 /luciferase plasmid described previously (pANK-1WT ; p296 from reference ) by using a quick ligation kit (New England Biolabs, Ipswich, MA). .. Maximum-efficiency DH5α bacteria (Invitrogen, Gaithersburg, MD) were transformed with the ligation products and grown for 16 h in LB medium containing ampicillin, after which plasmid DNA was prepared.

    Article Title: Herpes simplex viruses activate phospholipid scramblase to redistribute phosphatidylserines and Akt to the outer leaflet of the plasma membrane and promote viral entry
    Article Snippet: .. 79VB4 cells were co-transfected with HSV-2(G) DNA and DNA from an engineered plasmid (designated pEF6/5UL1HygroGFP) that contains the gL-2 gene (UL1) disrupted at the BmgBI and HpaI sites (R0628, R0105, New England BioLabs) by the hygromycin enhanced green fluorescent protein (EGFP) fusion protein under the control of the immediate early promoter of human cytomegalovirus (HCMV) constructed from pHygGFP (Clontech) (The plasmid encoding HSV-2(G) gL-2, designated pEF6/5 UL1 was a gift from Torin Weisbrod and William Jacobs).This dual function marker vector allows for drug selection with the ability to identify positive transfectants using GFP as a fluorescent reporter. .. Cotran sfections were performed using the Effectene Transfection reagent (301425, Qiagen).

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    New England Biolabs bmgb i
    PCR RFLP of rrs gene after digestion with <t>BmgB</t> I. Lane 1, 100 bp DNA ladder; Lanes 2 and 3, 353 and 128 bp fragments with wild-type sequence; and Lanes 4 and 5, 481 bp fragments of mutant sequence
    Bmgb I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bmgb i/product/New England Biolabs
    Average 94 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
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    PCR RFLP of rrs gene after digestion with BmgB I. Lane 1, 100 bp DNA ladder; Lanes 2 and 3, 353 and 128 bp fragments with wild-type sequence; and Lanes 4 and 5, 481 bp fragments of mutant sequence

    Journal: Journal of Epidemiology and Global Health

    Article Title: Genetic Characterization of Second-Line Drug-Resistant and Extensively Drug-Resistant Mycobacterium tuberculosis from the Northern Region of India

    doi: 10.2991/j.jegh.2018.02.100

    Figure Lengend Snippet: PCR RFLP of rrs gene after digestion with BmgB I. Lane 1, 100 bp DNA ladder; Lanes 2 and 3, 353 and 128 bp fragments with wild-type sequence; and Lanes 4 and 5, 481 bp fragments of mutant sequence

    Article Snippet: For PCR RFLP, the same primers RRSF and RRSR were used followed by the restriction digestion with BmgB I (New England Biolabs, UK).

    Techniques: Polymerase Chain Reaction, Sequencing, Mutagenesis