bsrgi hf  (New England Biolabs)


Bioz Verified Symbol New England Biolabs is a verified supplier
Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95
    Name:
    BsrGI HF
    Description:
    BsrGI HF 5 000 units
    Catalog Number:
    R3575L
    Price:
    282
    Category:
    Restriction Enzymes
    Size:
    5 000 units
    Buy from Supplier


    Structured Review

    New England Biolabs bsrgi hf
    BsrGI HF
    BsrGI HF 5 000 units
    https://www.bioz.com/result/bsrgi hf/product/New England Biolabs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bsrgi hf - by Bioz Stars, 2021-04
    95/100 stars

    Images

    Related Articles

    Plasmid Preparation:

    Article Title: Genome-scale CRISPR screening at high sensitivity with an empirically designed sgRNA library
    Article Snippet: Single sgRNA cloningSingle sgRNA sequences were cloned into the HDCRISPRv1 vector as described previously [ ]. .. In brief, the HDCRISPRv1 vector was sequentially digested with BfuAI (NEB) and BsrGI-HF (NEB), followed by dephosphorylation using CIP (NEB). .. The digested backbone was gel purified using the Macherey & Nagel NucleoSpin Gel and PCR Clean-up kit. sgRNA inserts were designed as two complementary oligos encoding the sgRNA target region as well as the cloning specific overhangs and were ordered as standard desalted oligos from Eurofins Genomics.

    Article Title: Dual RNA-Seq Unveils the Role of the Pseudomonas plecoglossicida fliA Gene in Pathogen-Host Interaction with Larimichthys crocea
    Article Snippet: Five siRNA sequences targeting the fliA gene were designed by BLOCK-iTTM RNAi Designer ( http://rnaidesigner.thermofisher.com/rnaiexpress/setOption.do?designOption=shrna & pid=708587103220684543 ) and synthetized by Shanghai Generay Biotech Co., Ltd. (Shanghai, China; ). .. First, the pCM130/tac vector was linearized using the restriction enzymes NsiI-HF and BsrGI-HF (New England Biolabs, Ipswich, MA, USA) and ligated to annealed shRNA with T4 DNA ligase. .. Second, the recombinant plasmid was transformed into E. coli DH5a by heat shock, extracted using EasyPure Plasmid MiniPrep Kit (TransGen Biotech, Beijing, China) and electroporated into P. plecoglossicida .

    Article Title: Pooled CRISPR screening at high sensitivity with an empirically designed sgRNA library
    Article Snippet: The resulting PCR product was purified using the NucleoSpin Gel and PCR Clean-up kit (Machery Nagel) and the correct fragment size was confirmed using a High Sensitivity Bioanalyzer DNA Kit (Agilent). .. For cloning, the HD CRISPR vector was digested with BfuAI (NEB) and BsrGI-HF (NEB) overnight and dephosphorylated using CIP (NEB). .. The resulting linearized ∼7000 bp vector missing the eGFP stuffer was gel purified again using the NucleoSpin Gel and PCR Clean-up kit (Machery Nagel) and used for library cloning in a one step digestion-ligation-reaction ( ).

    Article Title: Deep mutational scanning of S. pyogenes Cas9 reveals important functional domains
    Article Snippet: These primers along with G-Cas For and G-Cas Rev (for mammalian expression, using the bacterial expression plasmid as template to help distinguish mutated products from template) or EF For and SV40 For (for bacterial expression, using the mammalian expression plasmid as template to help distinguish mutated products from template) were used to generate the overlapping fragments necessary to produce the chosen mutation combination. .. The fused PCR products were either digested with BstXI and BsrGI-HF (NEB) and ligated back into the mammalian expression vector EF-SpCas9 (Supplementary Figure ) for mammalian expression or used for golden gate assembly into pUC-ProD-LacZ-T2 for bacterial expression as described above. .. All primers used for mutation PCRs can be found in Supplementary Table .

    Article Title: Identification of Mubritinib (TAK 165) as an inhibitor of KSHV driven primary effusion lymphoma via disruption of mitochondrial OXPHOS metabolism
    Article Snippet: Finally, complementary oligonucleotides containing the three LANA DNA binding sites (LBS2-LBS1-LBS3) were inserted upstream of the MLP promoter using AflII and NheI enzymes (New England Biolabs). .. The RMCE-HILO donor plasmid was generated by digesting the pEM791 plasmid [ ] with AgeI-HF and BsrGI-HF enzymes (New England Biolabs) and inserting two PCR amplified sequences containing the FLAG tag fused to the VP16 activation domain and the LANA DNA-binding domain by Gibson Assembly (New England Biolabs). .. Cell lines The RMCE-HILO HEK293T acceptor cells were obtained from Eugene V. Makeyev (Nanyang Technological University, Singapore) and maintained in Dulbecco’s modified Eagle’s medium (Gibco BRL) with 10% FetalPlex Serum Complex (Gemini Bio) and penicillin and streptomycin (50 U/ml).

    De-Phosphorylation Assay:

    Article Title: Genome-scale CRISPR screening at high sensitivity with an empirically designed sgRNA library
    Article Snippet: Single sgRNA cloningSingle sgRNA sequences were cloned into the HDCRISPRv1 vector as described previously [ ]. .. In brief, the HDCRISPRv1 vector was sequentially digested with BfuAI (NEB) and BsrGI-HF (NEB), followed by dephosphorylation using CIP (NEB). .. The digested backbone was gel purified using the Macherey & Nagel NucleoSpin Gel and PCR Clean-up kit. sgRNA inserts were designed as two complementary oligos encoding the sgRNA target region as well as the cloning specific overhangs and were ordered as standard desalted oligos from Eurofins Genomics.

    Amplification:

    Article Title: A genome-wide screen in the mouse liver reveals sex-specific and cell non-autonomous regulation of cell fitness
    Article Snippet: .. The amplicon and pLCv2-opti-stuffer-mCherry-puro (or -mTurq2-puro) were digested with BsrGI-HF and PmeI (New England Biolabs) and purified as above. .. NEBuilder HiFi DNA Assembly Master Mix (New England Biolabs) was used to assemble 25 ng each of vector and fragment in a 20 μL reaction for 15 min at 50 °C.

    Article Title: Identification of Mubritinib (TAK 165) as an inhibitor of KSHV driven primary effusion lymphoma via disruption of mitochondrial OXPHOS metabolism
    Article Snippet: Finally, complementary oligonucleotides containing the three LANA DNA binding sites (LBS2-LBS1-LBS3) were inserted upstream of the MLP promoter using AflII and NheI enzymes (New England Biolabs). .. The RMCE-HILO donor plasmid was generated by digesting the pEM791 plasmid [ ] with AgeI-HF and BsrGI-HF enzymes (New England Biolabs) and inserting two PCR amplified sequences containing the FLAG tag fused to the VP16 activation domain and the LANA DNA-binding domain by Gibson Assembly (New England Biolabs). .. Cell lines The RMCE-HILO HEK293T acceptor cells were obtained from Eugene V. Makeyev (Nanyang Technological University, Singapore) and maintained in Dulbecco’s modified Eagle’s medium (Gibco BRL) with 10% FetalPlex Serum Complex (Gemini Bio) and penicillin and streptomycin (50 U/ml).

    Purification:

    Article Title: A genome-wide screen in the mouse liver reveals sex-specific and cell non-autonomous regulation of cell fitness
    Article Snippet: .. The amplicon and pLCv2-opti-stuffer-mCherry-puro (or -mTurq2-puro) were digested with BsrGI-HF and PmeI (New England Biolabs) and purified as above. .. NEBuilder HiFi DNA Assembly Master Mix (New England Biolabs) was used to assemble 25 ng each of vector and fragment in a 20 μL reaction for 15 min at 50 °C.

    shRNA:

    Article Title: Dual RNA-Seq Unveils the Role of the Pseudomonas plecoglossicida fliA Gene in Pathogen-Host Interaction with Larimichthys crocea
    Article Snippet: Five siRNA sequences targeting the fliA gene were designed by BLOCK-iTTM RNAi Designer ( http://rnaidesigner.thermofisher.com/rnaiexpress/setOption.do?designOption=shrna & pid=708587103220684543 ) and synthetized by Shanghai Generay Biotech Co., Ltd. (Shanghai, China; ). .. First, the pCM130/tac vector was linearized using the restriction enzymes NsiI-HF and BsrGI-HF (New England Biolabs, Ipswich, MA, USA) and ligated to annealed shRNA with T4 DNA ligase. .. Second, the recombinant plasmid was transformed into E. coli DH5a by heat shock, extracted using EasyPure Plasmid MiniPrep Kit (TransGen Biotech, Beijing, China) and electroporated into P. plecoglossicida .

    Clone Assay:

    Article Title: Pooled CRISPR screening at high sensitivity with an empirically designed sgRNA library
    Article Snippet: The resulting PCR product was purified using the NucleoSpin Gel and PCR Clean-up kit (Machery Nagel) and the correct fragment size was confirmed using a High Sensitivity Bioanalyzer DNA Kit (Agilent). .. For cloning, the HD CRISPR vector was digested with BfuAI (NEB) and BsrGI-HF (NEB) overnight and dephosphorylated using CIP (NEB). .. The resulting linearized ∼7000 bp vector missing the eGFP stuffer was gel purified again using the NucleoSpin Gel and PCR Clean-up kit (Machery Nagel) and used for library cloning in a one step digestion-ligation-reaction ( ).

    CRISPR:

    Article Title: Pooled CRISPR screening at high sensitivity with an empirically designed sgRNA library
    Article Snippet: The resulting PCR product was purified using the NucleoSpin Gel and PCR Clean-up kit (Machery Nagel) and the correct fragment size was confirmed using a High Sensitivity Bioanalyzer DNA Kit (Agilent). .. For cloning, the HD CRISPR vector was digested with BfuAI (NEB) and BsrGI-HF (NEB) overnight and dephosphorylated using CIP (NEB). .. The resulting linearized ∼7000 bp vector missing the eGFP stuffer was gel purified again using the NucleoSpin Gel and PCR Clean-up kit (Machery Nagel) and used for library cloning in a one step digestion-ligation-reaction ( ).

    Polymerase Chain Reaction:

    Article Title: A versatile vector for mycobacterial protein production with a functional minimized acetamidase regulon
    Article Snippet: The linearized pMyCA was amplified by polymerase chain reaction (PCR) using the ΔamiD‐S‐BsrGI primer pair (Table ). .. The primer pair was designed with a BsrGI restriction recognition site (BsrGI‐HF, New England Biolabs® ) for the circularization of the linear PCR product. .. The plasmid pMyCA‐esxBA, which contains the coding sequence for the cfp10‐esat6 ( esxB/Rv3874 and esxA/Rv3875 ) operon from M. tuberculosis H37Rv was generated from pMyNT‐esxBA , using the same ΔamiDS‐BsrGI primer pair.

    Article Title: Deep mutational scanning of S. pyogenes Cas9 reveals important functional domains
    Article Snippet: These primers along with G-Cas For and G-Cas Rev (for mammalian expression, using the bacterial expression plasmid as template to help distinguish mutated products from template) or EF For and SV40 For (for bacterial expression, using the mammalian expression plasmid as template to help distinguish mutated products from template) were used to generate the overlapping fragments necessary to produce the chosen mutation combination. .. The fused PCR products were either digested with BstXI and BsrGI-HF (NEB) and ligated back into the mammalian expression vector EF-SpCas9 (Supplementary Figure ) for mammalian expression or used for golden gate assembly into pUC-ProD-LacZ-T2 for bacterial expression as described above. .. All primers used for mutation PCRs can be found in Supplementary Table .

    Article Title: Identification of Mubritinib (TAK 165) as an inhibitor of KSHV driven primary effusion lymphoma via disruption of mitochondrial OXPHOS metabolism
    Article Snippet: Finally, complementary oligonucleotides containing the three LANA DNA binding sites (LBS2-LBS1-LBS3) were inserted upstream of the MLP promoter using AflII and NheI enzymes (New England Biolabs). .. The RMCE-HILO donor plasmid was generated by digesting the pEM791 plasmid [ ] with AgeI-HF and BsrGI-HF enzymes (New England Biolabs) and inserting two PCR amplified sequences containing the FLAG tag fused to the VP16 activation domain and the LANA DNA-binding domain by Gibson Assembly (New England Biolabs). .. Cell lines The RMCE-HILO HEK293T acceptor cells were obtained from Eugene V. Makeyev (Nanyang Technological University, Singapore) and maintained in Dulbecco’s modified Eagle’s medium (Gibco BRL) with 10% FetalPlex Serum Complex (Gemini Bio) and penicillin and streptomycin (50 U/ml).

    Expressing:

    Article Title: Deep mutational scanning of S. pyogenes Cas9 reveals important functional domains
    Article Snippet: These primers along with G-Cas For and G-Cas Rev (for mammalian expression, using the bacterial expression plasmid as template to help distinguish mutated products from template) or EF For and SV40 For (for bacterial expression, using the mammalian expression plasmid as template to help distinguish mutated products from template) were used to generate the overlapping fragments necessary to produce the chosen mutation combination. .. The fused PCR products were either digested with BstXI and BsrGI-HF (NEB) and ligated back into the mammalian expression vector EF-SpCas9 (Supplementary Figure ) for mammalian expression or used for golden gate assembly into pUC-ProD-LacZ-T2 for bacterial expression as described above. .. All primers used for mutation PCRs can be found in Supplementary Table .

    Generated:

    Article Title: Identification of Mubritinib (TAK 165) as an inhibitor of KSHV driven primary effusion lymphoma via disruption of mitochondrial OXPHOS metabolism
    Article Snippet: Finally, complementary oligonucleotides containing the three LANA DNA binding sites (LBS2-LBS1-LBS3) were inserted upstream of the MLP promoter using AflII and NheI enzymes (New England Biolabs). .. The RMCE-HILO donor plasmid was generated by digesting the pEM791 plasmid [ ] with AgeI-HF and BsrGI-HF enzymes (New England Biolabs) and inserting two PCR amplified sequences containing the FLAG tag fused to the VP16 activation domain and the LANA DNA-binding domain by Gibson Assembly (New England Biolabs). .. Cell lines The RMCE-HILO HEK293T acceptor cells were obtained from Eugene V. Makeyev (Nanyang Technological University, Singapore) and maintained in Dulbecco’s modified Eagle’s medium (Gibco BRL) with 10% FetalPlex Serum Complex (Gemini Bio) and penicillin and streptomycin (50 U/ml).

    FLAG-tag:

    Article Title: Identification of Mubritinib (TAK 165) as an inhibitor of KSHV driven primary effusion lymphoma via disruption of mitochondrial OXPHOS metabolism
    Article Snippet: Finally, complementary oligonucleotides containing the three LANA DNA binding sites (LBS2-LBS1-LBS3) were inserted upstream of the MLP promoter using AflII and NheI enzymes (New England Biolabs). .. The RMCE-HILO donor plasmid was generated by digesting the pEM791 plasmid [ ] with AgeI-HF and BsrGI-HF enzymes (New England Biolabs) and inserting two PCR amplified sequences containing the FLAG tag fused to the VP16 activation domain and the LANA DNA-binding domain by Gibson Assembly (New England Biolabs). .. Cell lines The RMCE-HILO HEK293T acceptor cells were obtained from Eugene V. Makeyev (Nanyang Technological University, Singapore) and maintained in Dulbecco’s modified Eagle’s medium (Gibco BRL) with 10% FetalPlex Serum Complex (Gemini Bio) and penicillin and streptomycin (50 U/ml).

    Activation Assay:

    Article Title: Identification of Mubritinib (TAK 165) as an inhibitor of KSHV driven primary effusion lymphoma via disruption of mitochondrial OXPHOS metabolism
    Article Snippet: Finally, complementary oligonucleotides containing the three LANA DNA binding sites (LBS2-LBS1-LBS3) were inserted upstream of the MLP promoter using AflII and NheI enzymes (New England Biolabs). .. The RMCE-HILO donor plasmid was generated by digesting the pEM791 plasmid [ ] with AgeI-HF and BsrGI-HF enzymes (New England Biolabs) and inserting two PCR amplified sequences containing the FLAG tag fused to the VP16 activation domain and the LANA DNA-binding domain by Gibson Assembly (New England Biolabs). .. Cell lines The RMCE-HILO HEK293T acceptor cells were obtained from Eugene V. Makeyev (Nanyang Technological University, Singapore) and maintained in Dulbecco’s modified Eagle’s medium (Gibco BRL) with 10% FetalPlex Serum Complex (Gemini Bio) and penicillin and streptomycin (50 U/ml).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95
    New England Biolabs bsrgi hf
    Bsrgi Hf, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bsrgi hf/product/New England Biolabs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bsrgi hf - by Bioz Stars, 2021-04
    95/100 stars
      Buy from Supplier

    Image Search Results