fastdigest xbai  (Thermo Fisher)


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    Name:
    FastDigest XbaI
    Description:
    5 T ↓C T A G A 3 3 A G A T C ↑T 5 Thermo Scientific FastDigest XbaI restriction enzyme recognizes T CTAGA site and cuts best at 37°C in 5 15 minutes using universal FastDigest Buffer Thermo Scientific FastDigest XbaI is one of an advanced line of fast restriction enzymes that are all 100 active in the universal FastDigest and FastDigest Green reaction buffers The universal buffer allows rapid single double or multiple DNA digestion within 5 15 minutes eliminating any need for buffer change or subsequent DNA clean up steps See Reaction Conditions for FastDigest Enzymes for a table of enzyme activity heat inactivation and incubation times for this and other FastDigest restriction enzymes DNA modifying enzymes such as Klenow Fragment T4 DNA Ligase alkaline phosphatases and T4 DNA Polymerase all have 100 activity in FastDigest Buffer Therefore enzymes for downstream applications can be directly added to the FastDigest reaction mix For additional convenience FastDigest Green Buffer includes a density reagent and two tracking dyes for direct loading of digestion reaction products on gels Short incubation times and optimal composition of the universal FastDigest Buffer eliminate star activity effects Features• 100 activity of all FastDigest enzymes in the universal buffer• 100 buffer compatibility with downstream applications• Complete digestion in 5 15 minutes• Direct loading on gels• No star activity• 176 FastDigest enzymes availableApplications• Molecular cloning• Restriction site mapping• Genotyping• Southern Blot• Restriction fragment length polymorphism RFLP • SNP analysisNote The FastDigest Green Buffer offers the same high performance in DNA digestion and downstream applications as the colorless FastDigest Buffer For applications that require product analysis by fluorescence excitation e g concentration measurements in UV light the colorless FastDigest Buffer is recommended For methylation sensitivity refer to product specifications
    Catalog Number:
    fd0684
    Price:
    None
    Category:
    Proteins Enzymes Peptides
    Applications:
    Cloning|Restriction Enzyme Cloning
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    Structured Review

    Thermo Fisher fastdigest xbai
    5 T ↓C T A G A 3 3 A G A T C ↑T 5 Thermo Scientific FastDigest XbaI restriction enzyme recognizes T CTAGA site and cuts best at 37°C in 5 15 minutes using universal FastDigest Buffer Thermo Scientific FastDigest XbaI is one of an advanced line of fast restriction enzymes that are all 100 active in the universal FastDigest and FastDigest Green reaction buffers The universal buffer allows rapid single double or multiple DNA digestion within 5 15 minutes eliminating any need for buffer change or subsequent DNA clean up steps See Reaction Conditions for FastDigest Enzymes for a table of enzyme activity heat inactivation and incubation times for this and other FastDigest restriction enzymes DNA modifying enzymes such as Klenow Fragment T4 DNA Ligase alkaline phosphatases and T4 DNA Polymerase all have 100 activity in FastDigest Buffer Therefore enzymes for downstream applications can be directly added to the FastDigest reaction mix For additional convenience FastDigest Green Buffer includes a density reagent and two tracking dyes for direct loading of digestion reaction products on gels Short incubation times and optimal composition of the universal FastDigest Buffer eliminate star activity effects Features• 100 activity of all FastDigest enzymes in the universal buffer• 100 buffer compatibility with downstream applications• Complete digestion in 5 15 minutes• Direct loading on gels• No star activity• 176 FastDigest enzymes availableApplications• Molecular cloning• Restriction site mapping• Genotyping• Southern Blot• Restriction fragment length polymorphism RFLP • SNP analysisNote The FastDigest Green Buffer offers the same high performance in DNA digestion and downstream applications as the colorless FastDigest Buffer For applications that require product analysis by fluorescence excitation e g concentration measurements in UV light the colorless FastDigest Buffer is recommended For methylation sensitivity refer to product specifications
    https://www.bioz.com/result/fastdigest xbai/product/Thermo Fisher
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    fastdigest xbai - by Bioz Stars, 2021-03
    93/100 stars

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    Related Articles

    Electrophoresis:

    Article Title: Design, Construction, and Validation of Histone-Binding Effectors in vitro and in Cells
    Article Snippet: Candidate clones were grown in 3 mL of liquid LB and 100 μ g/mL ampicillin. .. Restriction digestion followed by electrophoresis (1% agarose in TAE) was used to analyze purified plasmid DNA (Sigma PLN350–1KT): 500−1000 ng of plasmid DNA, 1 μ L each of FastDigest enzymes Xba I and PstI, and 1× FastDigest buffer (Thermo Fisher FD0684, FD0614, and B72). .. HEK293 Gal4-EED/luc cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% tetracycline-free fetal bovine serum and 1% penicillin and streptomycin (pen/strep) at 37 °C in a humidified CO2 incubator.

    Purification:

    Article Title: Design, Construction, and Validation of Histone-Binding Effectors in vitro and in Cells
    Article Snippet: Candidate clones were grown in 3 mL of liquid LB and 100 μ g/mL ampicillin. .. Restriction digestion followed by electrophoresis (1% agarose in TAE) was used to analyze purified plasmid DNA (Sigma PLN350–1KT): 500−1000 ng of plasmid DNA, 1 μ L each of FastDigest enzymes Xba I and PstI, and 1× FastDigest buffer (Thermo Fisher FD0684, FD0614, and B72). .. HEK293 Gal4-EED/luc cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% tetracycline-free fetal bovine serum and 1% penicillin and streptomycin (pen/strep) at 37 °C in a humidified CO2 incubator.

    Plasmid Preparation:

    Article Title: Design, Construction, and Validation of Histone-Binding Effectors in vitro and in Cells
    Article Snippet: Candidate clones were grown in 3 mL of liquid LB and 100 μ g/mL ampicillin. .. Restriction digestion followed by electrophoresis (1% agarose in TAE) was used to analyze purified plasmid DNA (Sigma PLN350–1KT): 500−1000 ng of plasmid DNA, 1 μ L each of FastDigest enzymes Xba I and PstI, and 1× FastDigest buffer (Thermo Fisher FD0684, FD0614, and B72). .. HEK293 Gal4-EED/luc cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% tetracycline-free fetal bovine serum and 1% penicillin and streptomycin (pen/strep) at 37 °C in a humidified CO2 incubator.

    Article Title: Amdinocillin (Mecillinam) Resistance Mutations in Clinical Isolates and Laboratory-Selected Mutants of Escherichia coli
    Article Snippet: Briefly, the insert ( E. coli MG1655 cysB gene with the native promoter and terminator) was PCR amplified with primers containing XbaI and PstI restriction enzyme cleavage sites, respectively. .. The insert and the plasmid were first cleaved with Fast Digest XbaI and PstI according to the manual provided by Thermo Fisher and then ligated with Ready-To-Go T4 DNA ligase (GE Health Care Life Sciences). .. The resulting plasmids were transformed into MG1655 by electroporation and then spread on Cam-supplemented MH agar plates for selection of plasmid uptake.

    Article Title: Characterisation of three fungal glucuronoyl esterases on glucuronic acid ester model compounds
    Article Snippet: .. Cloning of putative fungal GE genes Codon-optimised genes excluding signal peptide were synthesised (NZYTech, Portugal) and cloned into the Pichia expression vector pPICZαA (Thermo Fisher Scientific) using FastDigest Eco RI and FastDigest Xba I (Thermo Fisher Scientific) in frame with the N-terminal α-factor and the C-terminal myc- and 6XHis-tag. .. Briefly, DNA molecules were purified using the Illustra GFX GelBand purification kit (GE Healthcare) and vector and respective gene inserts ligated with T4 DNA ligase according to the manufacturer’s protocol (Thermo Fisher Scientific).

    Article Title: Design, Construction, and Validation of Histone-Binding Effectors in vitro and in Cells
    Article Snippet: We recommend the following colony PCR protocol to quickly screen for forward insertions. (1) Assemble constructs via nondirectional cloning of fusion-coding regions into vector MV10 using restriction digests and ligations as previously described. (2) Prepare colony PCR mixes as follows: 1× GoTaq green master mix (Promega M7122), 0.4 μ M forward primer 5′-caccatcgtggaacagtacg, and 0.4 μ M reverse primer 5′-gcaactagaaggcacagtcg in a final volume of 25 μ L in each 0.5 mL tube. (3) Pick colonies grown from ligation-transformed E. coli with a sterile, disposable micropipette tip, streak onto a small area (1 cm2 ) on a gridded and labeled LB agar, 100 μ g/mL ampicillin plate, swirl the same tip in each PCR mix, and discard the tip. .. Incubate the streak plate overnight at 37 °C. (4) Perform PCR as follows: 95 °C for 2 min; 25 cycles of 95 °C for 30 s, 55 °C for 30 s, and 72 °C for 1 min; 72 °C for 5 min; and held at 4 °C. (5) Analyze 20−50% of each reaction mixture via gel electrophoresis. (6) Inoculate 5 mL of liquid LB, 100 μ g/mL ampicillin cultures with candidate clones from the streak plate, grow for 18 h at 37 °C with shaking, and miniprep the plasmid DNA (Sigma PLN350–1KT). (7) Perform restriction digests as follows: 500−1000 ng of plasmid DNA, 1 μ L of each of FastDigest enzymes XbaI and Pst I, and 1× FastDigest buffer (Thermo Fisher FD0684, FD0614, and B72). .. Analyze the products using standard agarose gel electrophoresis with TAE buffer.

    Article Title: Transcriptional responses to IFN-γ require Mediator kinase-dependent pause release and mechanistically distinct functions of CDK8 and CDK19
    Article Snippet: Primers of the second PCR were designed in a way that they flank the CDS, remove the stop codon and introduce a 5’ XbaI and 3’ XhoI NotI cleavage site (Forward: 5’-CATTCTAGACCGAGGAGTCCCTTGCTGAA-3’, Reverse: 5’- TATGCGGCCGCTATCTCGAGTACCGGTGGGTCTGGTGAGAT-3’). .. After gel purification and sequence validation, the PCR product and the PiggyBac-EF1-MCS-IRES-Neo cDNA Cloning and Expression Vector (#PB533A-2, SBI System Biosciences) were double digested using XbaI (#FD0684, Thermo Scientific) and NotI (#FD0594, Thermo Scientific) followed by gel purification, T4 DNA Ligase (#EL0011, Thermo Scientific) mediated ligation, transformation into chemically competent DH10B and ampicillin selection (100 pg/ml, #A0839, AppliChem). .. Colonies were PCR-screened (Forward: 5’- CAATTGAACGGGTGCCTAGAG-3’, Reverse: 5’-CCTTGTTGAATACGCTTGAGGAGA-3’) and plasmids were isolated by using plasmid mini prep kit (QIAprep, #27106, Qiagen).

    Clone Assay:

    Article Title: Characterisation of three fungal glucuronoyl esterases on glucuronic acid ester model compounds
    Article Snippet: .. Cloning of putative fungal GE genes Codon-optimised genes excluding signal peptide were synthesised (NZYTech, Portugal) and cloned into the Pichia expression vector pPICZαA (Thermo Fisher Scientific) using FastDigest Eco RI and FastDigest Xba I (Thermo Fisher Scientific) in frame with the N-terminal α-factor and the C-terminal myc- and 6XHis-tag. .. Briefly, DNA molecules were purified using the Illustra GFX GelBand purification kit (GE Healthcare) and vector and respective gene inserts ligated with T4 DNA ligase according to the manufacturer’s protocol (Thermo Fisher Scientific).

    Article Title: Design, Construction, and Validation of Histone-Binding Effectors in vitro and in Cells
    Article Snippet: We recommend the following colony PCR protocol to quickly screen for forward insertions. (1) Assemble constructs via nondirectional cloning of fusion-coding regions into vector MV10 using restriction digests and ligations as previously described. (2) Prepare colony PCR mixes as follows: 1× GoTaq green master mix (Promega M7122), 0.4 μ M forward primer 5′-caccatcgtggaacagtacg, and 0.4 μ M reverse primer 5′-gcaactagaaggcacagtcg in a final volume of 25 μ L in each 0.5 mL tube. (3) Pick colonies grown from ligation-transformed E. coli with a sterile, disposable micropipette tip, streak onto a small area (1 cm2 ) on a gridded and labeled LB agar, 100 μ g/mL ampicillin plate, swirl the same tip in each PCR mix, and discard the tip. .. Incubate the streak plate overnight at 37 °C. (4) Perform PCR as follows: 95 °C for 2 min; 25 cycles of 95 °C for 30 s, 55 °C for 30 s, and 72 °C for 1 min; 72 °C for 5 min; and held at 4 °C. (5) Analyze 20−50% of each reaction mixture via gel electrophoresis. (6) Inoculate 5 mL of liquid LB, 100 μ g/mL ampicillin cultures with candidate clones from the streak plate, grow for 18 h at 37 °C with shaking, and miniprep the plasmid DNA (Sigma PLN350–1KT). (7) Perform restriction digests as follows: 500−1000 ng of plasmid DNA, 1 μ L of each of FastDigest enzymes XbaI and Pst I, and 1× FastDigest buffer (Thermo Fisher FD0684, FD0614, and B72). .. Analyze the products using standard agarose gel electrophoresis with TAE buffer.

    Article Title: Transcriptional responses to IFN-γ require Mediator kinase-dependent pause release and mechanistically distinct functions of CDK8 and CDK19
    Article Snippet: Primers of the second PCR were designed in a way that they flank the CDS, remove the stop codon and introduce a 5’ XbaI and 3’ XhoI NotI cleavage site (Forward: 5’-CATTCTAGACCGAGGAGTCCCTTGCTGAA-3’, Reverse: 5’- TATGCGGCCGCTATCTCGAGTACCGGTGGGTCTGGTGAGAT-3’). .. After gel purification and sequence validation, the PCR product and the PiggyBac-EF1-MCS-IRES-Neo cDNA Cloning and Expression Vector (#PB533A-2, SBI System Biosciences) were double digested using XbaI (#FD0684, Thermo Scientific) and NotI (#FD0594, Thermo Scientific) followed by gel purification, T4 DNA Ligase (#EL0011, Thermo Scientific) mediated ligation, transformation into chemically competent DH10B and ampicillin selection (100 pg/ml, #A0839, AppliChem). .. Colonies were PCR-screened (Forward: 5’- CAATTGAACGGGTGCCTAGAG-3’, Reverse: 5’-CCTTGTTGAATACGCTTGAGGAGA-3’) and plasmids were isolated by using plasmid mini prep kit (QIAprep, #27106, Qiagen).

    Expressing:

    Article Title: Characterisation of three fungal glucuronoyl esterases on glucuronic acid ester model compounds
    Article Snippet: .. Cloning of putative fungal GE genes Codon-optimised genes excluding signal peptide were synthesised (NZYTech, Portugal) and cloned into the Pichia expression vector pPICZαA (Thermo Fisher Scientific) using FastDigest Eco RI and FastDigest Xba I (Thermo Fisher Scientific) in frame with the N-terminal α-factor and the C-terminal myc- and 6XHis-tag. .. Briefly, DNA molecules were purified using the Illustra GFX GelBand purification kit (GE Healthcare) and vector and respective gene inserts ligated with T4 DNA ligase according to the manufacturer’s protocol (Thermo Fisher Scientific).

    Article Title: Transcriptional responses to IFN-γ require Mediator kinase-dependent pause release and mechanistically distinct functions of CDK8 and CDK19
    Article Snippet: Primers of the second PCR were designed in a way that they flank the CDS, remove the stop codon and introduce a 5’ XbaI and 3’ XhoI NotI cleavage site (Forward: 5’-CATTCTAGACCGAGGAGTCCCTTGCTGAA-3’, Reverse: 5’- TATGCGGCCGCTATCTCGAGTACCGGTGGGTCTGGTGAGAT-3’). .. After gel purification and sequence validation, the PCR product and the PiggyBac-EF1-MCS-IRES-Neo cDNA Cloning and Expression Vector (#PB533A-2, SBI System Biosciences) were double digested using XbaI (#FD0684, Thermo Scientific) and NotI (#FD0594, Thermo Scientific) followed by gel purification, T4 DNA Ligase (#EL0011, Thermo Scientific) mediated ligation, transformation into chemically competent DH10B and ampicillin selection (100 pg/ml, #A0839, AppliChem). .. Colonies were PCR-screened (Forward: 5’- CAATTGAACGGGTGCCTAGAG-3’, Reverse: 5’-CCTTGTTGAATACGCTTGAGGAGA-3’) and plasmids were isolated by using plasmid mini prep kit (QIAprep, #27106, Qiagen).

    Polymerase Chain Reaction:

    Article Title: Design, Construction, and Validation of Histone-Binding Effectors in vitro and in Cells
    Article Snippet: We recommend the following colony PCR protocol to quickly screen for forward insertions. (1) Assemble constructs via nondirectional cloning of fusion-coding regions into vector MV10 using restriction digests and ligations as previously described. (2) Prepare colony PCR mixes as follows: 1× GoTaq green master mix (Promega M7122), 0.4 μ M forward primer 5′-caccatcgtggaacagtacg, and 0.4 μ M reverse primer 5′-gcaactagaaggcacagtcg in a final volume of 25 μ L in each 0.5 mL tube. (3) Pick colonies grown from ligation-transformed E. coli with a sterile, disposable micropipette tip, streak onto a small area (1 cm2 ) on a gridded and labeled LB agar, 100 μ g/mL ampicillin plate, swirl the same tip in each PCR mix, and discard the tip. .. Incubate the streak plate overnight at 37 °C. (4) Perform PCR as follows: 95 °C for 2 min; 25 cycles of 95 °C for 30 s, 55 °C for 30 s, and 72 °C for 1 min; 72 °C for 5 min; and held at 4 °C. (5) Analyze 20−50% of each reaction mixture via gel electrophoresis. (6) Inoculate 5 mL of liquid LB, 100 μ g/mL ampicillin cultures with candidate clones from the streak plate, grow for 18 h at 37 °C with shaking, and miniprep the plasmid DNA (Sigma PLN350–1KT). (7) Perform restriction digests as follows: 500−1000 ng of plasmid DNA, 1 μ L of each of FastDigest enzymes XbaI and Pst I, and 1× FastDigest buffer (Thermo Fisher FD0684, FD0614, and B72). .. Analyze the products using standard agarose gel electrophoresis with TAE buffer.

    Article Title: Transcriptional responses to IFN-γ require Mediator kinase-dependent pause release and mechanistically distinct functions of CDK8 and CDK19
    Article Snippet: Primers of the second PCR were designed in a way that they flank the CDS, remove the stop codon and introduce a 5’ XbaI and 3’ XhoI NotI cleavage site (Forward: 5’-CATTCTAGACCGAGGAGTCCCTTGCTGAA-3’, Reverse: 5’- TATGCGGCCGCTATCTCGAGTACCGGTGGGTCTGGTGAGAT-3’). .. After gel purification and sequence validation, the PCR product and the PiggyBac-EF1-MCS-IRES-Neo cDNA Cloning and Expression Vector (#PB533A-2, SBI System Biosciences) were double digested using XbaI (#FD0684, Thermo Scientific) and NotI (#FD0594, Thermo Scientific) followed by gel purification, T4 DNA Ligase (#EL0011, Thermo Scientific) mediated ligation, transformation into chemically competent DH10B and ampicillin selection (100 pg/ml, #A0839, AppliChem). .. Colonies were PCR-screened (Forward: 5’- CAATTGAACGGGTGCCTAGAG-3’, Reverse: 5’-CCTTGTTGAATACGCTTGAGGAGA-3’) and plasmids were isolated by using plasmid mini prep kit (QIAprep, #27106, Qiagen).

    Nucleic Acid Electrophoresis:

    Article Title: Design, Construction, and Validation of Histone-Binding Effectors in vitro and in Cells
    Article Snippet: We recommend the following colony PCR protocol to quickly screen for forward insertions. (1) Assemble constructs via nondirectional cloning of fusion-coding regions into vector MV10 using restriction digests and ligations as previously described. (2) Prepare colony PCR mixes as follows: 1× GoTaq green master mix (Promega M7122), 0.4 μ M forward primer 5′-caccatcgtggaacagtacg, and 0.4 μ M reverse primer 5′-gcaactagaaggcacagtcg in a final volume of 25 μ L in each 0.5 mL tube. (3) Pick colonies grown from ligation-transformed E. coli with a sterile, disposable micropipette tip, streak onto a small area (1 cm2 ) on a gridded and labeled LB agar, 100 μ g/mL ampicillin plate, swirl the same tip in each PCR mix, and discard the tip. .. Incubate the streak plate overnight at 37 °C. (4) Perform PCR as follows: 95 °C for 2 min; 25 cycles of 95 °C for 30 s, 55 °C for 30 s, and 72 °C for 1 min; 72 °C for 5 min; and held at 4 °C. (5) Analyze 20−50% of each reaction mixture via gel electrophoresis. (6) Inoculate 5 mL of liquid LB, 100 μ g/mL ampicillin cultures with candidate clones from the streak plate, grow for 18 h at 37 °C with shaking, and miniprep the plasmid DNA (Sigma PLN350–1KT). (7) Perform restriction digests as follows: 500−1000 ng of plasmid DNA, 1 μ L of each of FastDigest enzymes XbaI and Pst I, and 1× FastDigest buffer (Thermo Fisher FD0684, FD0614, and B72). .. Analyze the products using standard agarose gel electrophoresis with TAE buffer.

    Gel Purification:

    Article Title: Transcriptional responses to IFN-γ require Mediator kinase-dependent pause release and mechanistically distinct functions of CDK8 and CDK19
    Article Snippet: Primers of the second PCR were designed in a way that they flank the CDS, remove the stop codon and introduce a 5’ XbaI and 3’ XhoI NotI cleavage site (Forward: 5’-CATTCTAGACCGAGGAGTCCCTTGCTGAA-3’, Reverse: 5’- TATGCGGCCGCTATCTCGAGTACCGGTGGGTCTGGTGAGAT-3’). .. After gel purification and sequence validation, the PCR product and the PiggyBac-EF1-MCS-IRES-Neo cDNA Cloning and Expression Vector (#PB533A-2, SBI System Biosciences) were double digested using XbaI (#FD0684, Thermo Scientific) and NotI (#FD0594, Thermo Scientific) followed by gel purification, T4 DNA Ligase (#EL0011, Thermo Scientific) mediated ligation, transformation into chemically competent DH10B and ampicillin selection (100 pg/ml, #A0839, AppliChem). .. Colonies were PCR-screened (Forward: 5’- CAATTGAACGGGTGCCTAGAG-3’, Reverse: 5’-CCTTGTTGAATACGCTTGAGGAGA-3’) and plasmids were isolated by using plasmid mini prep kit (QIAprep, #27106, Qiagen).

    Sequencing:

    Article Title: Transcriptional responses to IFN-γ require Mediator kinase-dependent pause release and mechanistically distinct functions of CDK8 and CDK19
    Article Snippet: Primers of the second PCR were designed in a way that they flank the CDS, remove the stop codon and introduce a 5’ XbaI and 3’ XhoI NotI cleavage site (Forward: 5’-CATTCTAGACCGAGGAGTCCCTTGCTGAA-3’, Reverse: 5’- TATGCGGCCGCTATCTCGAGTACCGGTGGGTCTGGTGAGAT-3’). .. After gel purification and sequence validation, the PCR product and the PiggyBac-EF1-MCS-IRES-Neo cDNA Cloning and Expression Vector (#PB533A-2, SBI System Biosciences) were double digested using XbaI (#FD0684, Thermo Scientific) and NotI (#FD0594, Thermo Scientific) followed by gel purification, T4 DNA Ligase (#EL0011, Thermo Scientific) mediated ligation, transformation into chemically competent DH10B and ampicillin selection (100 pg/ml, #A0839, AppliChem). .. Colonies were PCR-screened (Forward: 5’- CAATTGAACGGGTGCCTAGAG-3’, Reverse: 5’-CCTTGTTGAATACGCTTGAGGAGA-3’) and plasmids were isolated by using plasmid mini prep kit (QIAprep, #27106, Qiagen).

    Ligation:

    Article Title: Transcriptional responses to IFN-γ require Mediator kinase-dependent pause release and mechanistically distinct functions of CDK8 and CDK19
    Article Snippet: Primers of the second PCR were designed in a way that they flank the CDS, remove the stop codon and introduce a 5’ XbaI and 3’ XhoI NotI cleavage site (Forward: 5’-CATTCTAGACCGAGGAGTCCCTTGCTGAA-3’, Reverse: 5’- TATGCGGCCGCTATCTCGAGTACCGGTGGGTCTGGTGAGAT-3’). .. After gel purification and sequence validation, the PCR product and the PiggyBac-EF1-MCS-IRES-Neo cDNA Cloning and Expression Vector (#PB533A-2, SBI System Biosciences) were double digested using XbaI (#FD0684, Thermo Scientific) and NotI (#FD0594, Thermo Scientific) followed by gel purification, T4 DNA Ligase (#EL0011, Thermo Scientific) mediated ligation, transformation into chemically competent DH10B and ampicillin selection (100 pg/ml, #A0839, AppliChem). .. Colonies were PCR-screened (Forward: 5’- CAATTGAACGGGTGCCTAGAG-3’, Reverse: 5’-CCTTGTTGAATACGCTTGAGGAGA-3’) and plasmids were isolated by using plasmid mini prep kit (QIAprep, #27106, Qiagen).

    Transformation Assay:

    Article Title: Transcriptional responses to IFN-γ require Mediator kinase-dependent pause release and mechanistically distinct functions of CDK8 and CDK19
    Article Snippet: Primers of the second PCR were designed in a way that they flank the CDS, remove the stop codon and introduce a 5’ XbaI and 3’ XhoI NotI cleavage site (Forward: 5’-CATTCTAGACCGAGGAGTCCCTTGCTGAA-3’, Reverse: 5’- TATGCGGCCGCTATCTCGAGTACCGGTGGGTCTGGTGAGAT-3’). .. After gel purification and sequence validation, the PCR product and the PiggyBac-EF1-MCS-IRES-Neo cDNA Cloning and Expression Vector (#PB533A-2, SBI System Biosciences) were double digested using XbaI (#FD0684, Thermo Scientific) and NotI (#FD0594, Thermo Scientific) followed by gel purification, T4 DNA Ligase (#EL0011, Thermo Scientific) mediated ligation, transformation into chemically competent DH10B and ampicillin selection (100 pg/ml, #A0839, AppliChem). .. Colonies were PCR-screened (Forward: 5’- CAATTGAACGGGTGCCTAGAG-3’, Reverse: 5’-CCTTGTTGAATACGCTTGAGGAGA-3’) and plasmids were isolated by using plasmid mini prep kit (QIAprep, #27106, Qiagen).

    Selection:

    Article Title: Transcriptional responses to IFN-γ require Mediator kinase-dependent pause release and mechanistically distinct functions of CDK8 and CDK19
    Article Snippet: Primers of the second PCR were designed in a way that they flank the CDS, remove the stop codon and introduce a 5’ XbaI and 3’ XhoI NotI cleavage site (Forward: 5’-CATTCTAGACCGAGGAGTCCCTTGCTGAA-3’, Reverse: 5’- TATGCGGCCGCTATCTCGAGTACCGGTGGGTCTGGTGAGAT-3’). .. After gel purification and sequence validation, the PCR product and the PiggyBac-EF1-MCS-IRES-Neo cDNA Cloning and Expression Vector (#PB533A-2, SBI System Biosciences) were double digested using XbaI (#FD0684, Thermo Scientific) and NotI (#FD0594, Thermo Scientific) followed by gel purification, T4 DNA Ligase (#EL0011, Thermo Scientific) mediated ligation, transformation into chemically competent DH10B and ampicillin selection (100 pg/ml, #A0839, AppliChem). .. Colonies were PCR-screened (Forward: 5’- CAATTGAACGGGTGCCTAGAG-3’, Reverse: 5’-CCTTGTTGAATACGCTTGAGGAGA-3’) and plasmids were isolated by using plasmid mini prep kit (QIAprep, #27106, Qiagen).

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  • 94
    Thermo Fisher fastdigest xbai
    Fastdigest Xbai, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fastdigest xbai/product/Thermo Fisher
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    fastdigest xbai - by Bioz Stars, 2021-03
    94/100 stars
      Buy from Supplier

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