apai  (Thermo Fisher)


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    Name:
    FastDigest ApaI
    Description:
    5 G G G C C ↓C 3 3 C ↑C C G G G 5 Thermo Scientific FastDigest ApaI restriction enzyme recognizes GGGCC C site and cuts best at 37°C in 5 15 minutes using universal FastDigest Buffer Isoschizomers PspOMI Thermo Scientific FastDigest ApaI 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:
    fd1414
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    Applications:
    Cloning|Restriction Enzyme Cloning
    Category:
    Proteins Enzymes Peptides
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    Structured Review

    Thermo Fisher apai
    Structural- and chemical probing of <t>DNA</t> and DNA-PNA complex formation in pMP178(75 repeats). A) Non-denaturing PAGE of DNA fragments mapped by BQQ-OP cleavage and chloroacetaldehyde (CAA) modification followed by primer extension (PE). Plasmid pMP178(75 repeats), was incubated in the absence (No PNA) or the presence of 10 μM of PNA (GAA-PNA or CTT-PNA) in buffer (10 mM sodium cacodylate, 100 mM NaCl, 2 mM MgCl 2 , pH 7.5). The plasmid was then chemically modified using 2% CAA or cleaved using 1 μM BQQ-OP. Non-treated plasmid was used in a set of four different controls (C) of the PE reactions C1= plasmid incubated in the absence of PNA. C2= plasmid incubated in the presence of GAA-PNA. C3= plasmid incubated in the presence of CTT-PNA. C4= plasmid not incubated. All samples were linearized by <t>ApaI</t> then subjected as templates for the PE reaction. Sequence ladders using dideoxynucleotides (C=ddCTP, T=ddTTP, G=ddGTP and A=ddATP), linearized pMP178(75 repeats) and the PE reaction control (C4) are shown as references. The DNA fragments detected in a PE reaction using the GAA (R)-strand as template, and the right gel panel shows the DNA fragments detected in a PE reaction using the CTT (Y)-strand as template. The A 1 to A 9 nucleotides flanking the repeats of the R-strand and T 1 to T 12 flanking in the Y-strand and the middle point of the repeat sequence are indicated (M). B) Models showing the most predominant DNA and DNA-PNA structures formed at pMP178(75 repeats) in the absence or the presence of CTT-PNA or GAA-PNA respectively. They correspond to: 1. 5′3′3′-pyrimidine motif H-DNA, 2. CTT-PNA stabilized 5′3′3′-pyrimidine motif H-DNA, 3. Triplex-invasion, 4. Intermolecular triplex and 5. Duplex-invasion structure. Thin line= purine strand, thick line= pyrimidine strand, grey line= PNA. Regions modified by CAA (π) or cleaved by BQQ-OP (o) are indicated.
    5 G G G C C ↓C 3 3 C ↑C C G G G 5 Thermo Scientific FastDigest ApaI restriction enzyme recognizes GGGCC C site and cuts best at 37°C in 5 15 minutes using universal FastDigest Buffer Isoschizomers PspOMI Thermo Scientific FastDigest ApaI 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/apai/product/Thermo Fisher
    Average 95 stars, based on 48 article reviews
    Price from $9.99 to $1999.99
    apai - by Bioz Stars, 2020-05
    95/100 stars

    Images

    1) Product Images from "Disruption of Higher Order DNA Structures in Friedreich’s Ataxia (GAA)n Repeats by PNA or LNA Targeting"

    Article Title: Disruption of Higher Order DNA Structures in Friedreich’s Ataxia (GAA)n Repeats by PNA or LNA Targeting

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0165788

    Structural- and chemical probing of DNA and DNA-PNA complex formation in pMP178(75 repeats). A) Non-denaturing PAGE of DNA fragments mapped by BQQ-OP cleavage and chloroacetaldehyde (CAA) modification followed by primer extension (PE). Plasmid pMP178(75 repeats), was incubated in the absence (No PNA) or the presence of 10 μM of PNA (GAA-PNA or CTT-PNA) in buffer (10 mM sodium cacodylate, 100 mM NaCl, 2 mM MgCl 2 , pH 7.5). The plasmid was then chemically modified using 2% CAA or cleaved using 1 μM BQQ-OP. Non-treated plasmid was used in a set of four different controls (C) of the PE reactions C1= plasmid incubated in the absence of PNA. C2= plasmid incubated in the presence of GAA-PNA. C3= plasmid incubated in the presence of CTT-PNA. C4= plasmid not incubated. All samples were linearized by ApaI then subjected as templates for the PE reaction. Sequence ladders using dideoxynucleotides (C=ddCTP, T=ddTTP, G=ddGTP and A=ddATP), linearized pMP178(75 repeats) and the PE reaction control (C4) are shown as references. The DNA fragments detected in a PE reaction using the GAA (R)-strand as template, and the right gel panel shows the DNA fragments detected in a PE reaction using the CTT (Y)-strand as template. The A 1 to A 9 nucleotides flanking the repeats of the R-strand and T 1 to T 12 flanking in the Y-strand and the middle point of the repeat sequence are indicated (M). B) Models showing the most predominant DNA and DNA-PNA structures formed at pMP178(75 repeats) in the absence or the presence of CTT-PNA or GAA-PNA respectively. They correspond to: 1. 5′3′3′-pyrimidine motif H-DNA, 2. CTT-PNA stabilized 5′3′3′-pyrimidine motif H-DNA, 3. Triplex-invasion, 4. Intermolecular triplex and 5. Duplex-invasion structure. Thin line= purine strand, thick line= pyrimidine strand, grey line= PNA. Regions modified by CAA (π) or cleaved by BQQ-OP (o) are indicated.
    Figure Legend Snippet: Structural- and chemical probing of DNA and DNA-PNA complex formation in pMP178(75 repeats). A) Non-denaturing PAGE of DNA fragments mapped by BQQ-OP cleavage and chloroacetaldehyde (CAA) modification followed by primer extension (PE). Plasmid pMP178(75 repeats), was incubated in the absence (No PNA) or the presence of 10 μM of PNA (GAA-PNA or CTT-PNA) in buffer (10 mM sodium cacodylate, 100 mM NaCl, 2 mM MgCl 2 , pH 7.5). The plasmid was then chemically modified using 2% CAA or cleaved using 1 μM BQQ-OP. Non-treated plasmid was used in a set of four different controls (C) of the PE reactions C1= plasmid incubated in the absence of PNA. C2= plasmid incubated in the presence of GAA-PNA. C3= plasmid incubated in the presence of CTT-PNA. C4= plasmid not incubated. All samples were linearized by ApaI then subjected as templates for the PE reaction. Sequence ladders using dideoxynucleotides (C=ddCTP, T=ddTTP, G=ddGTP and A=ddATP), linearized pMP178(75 repeats) and the PE reaction control (C4) are shown as references. The DNA fragments detected in a PE reaction using the GAA (R)-strand as template, and the right gel panel shows the DNA fragments detected in a PE reaction using the CTT (Y)-strand as template. The A 1 to A 9 nucleotides flanking the repeats of the R-strand and T 1 to T 12 flanking in the Y-strand and the middle point of the repeat sequence are indicated (M). B) Models showing the most predominant DNA and DNA-PNA structures formed at pMP178(75 repeats) in the absence or the presence of CTT-PNA or GAA-PNA respectively. They correspond to: 1. 5′3′3′-pyrimidine motif H-DNA, 2. CTT-PNA stabilized 5′3′3′-pyrimidine motif H-DNA, 3. Triplex-invasion, 4. Intermolecular triplex and 5. Duplex-invasion structure. Thin line= purine strand, thick line= pyrimidine strand, grey line= PNA. Regions modified by CAA (π) or cleaved by BQQ-OP (o) are indicated.

    Techniques Used: Polyacrylamide Gel Electrophoresis, Cellular Antioxidant Activity Assay, Modification, Plasmid Preparation, Incubation, Sequencing

    BQQ-OP mediated DNA cleavage of H-DNA forming (GAA) 115 repeats in the presence of PNA. A) Schematic presentation of pMP179 and the H-DNA forming site. The two DNA fragments generated by triplex-specific cleavage followed by enzymatic digestion are indicated as X (3814 bp) and Y (3178 bp). B) Representative gel for pMP179(115 repeats) incubated with 10 μM PNA (CTT-PNA or GAA-PNA, lane 1 or 2, respectively), in the absence of ONs (lane 3) or with 10 μM ssDNA (CTT-DNA or GAA-DNA, lane 4 or 5, respectively) in buffer (10 mM sodium cacodylate, 100 mM NaCl, 2 mM MgCl 2 , pH 7.5). Triplex-specific cleavage was carried out in the presence of Cu 2+ and MPA, followed by digestion with ApaI. Reference supercoiled (SC) and linearized (Lin) pMP179(115 repeats) and a molecular weight DNA ladder (M) are also shown. C) Graph represents the percentage of BQQ-OP mediated cleavage obtained for pMP179(115 repeats) in the absence (indicate in the graph as pMP179) or in the presence of PNA- or DNA oligomers. Values indicate the ratio between the intensity of DNA double strand cleavage to the total band intensity from the respective lane and are expressed as mean±S.D. (n=3). No cleavage was obtained in the presence of GAA-PNA and therefore not included in the graph. ** P ≤0.01 in relation to plasmid in the absence of oligonucleotide (pMP179).
    Figure Legend Snippet: BQQ-OP mediated DNA cleavage of H-DNA forming (GAA) 115 repeats in the presence of PNA. A) Schematic presentation of pMP179 and the H-DNA forming site. The two DNA fragments generated by triplex-specific cleavage followed by enzymatic digestion are indicated as X (3814 bp) and Y (3178 bp). B) Representative gel for pMP179(115 repeats) incubated with 10 μM PNA (CTT-PNA or GAA-PNA, lane 1 or 2, respectively), in the absence of ONs (lane 3) or with 10 μM ssDNA (CTT-DNA or GAA-DNA, lane 4 or 5, respectively) in buffer (10 mM sodium cacodylate, 100 mM NaCl, 2 mM MgCl 2 , pH 7.5). Triplex-specific cleavage was carried out in the presence of Cu 2+ and MPA, followed by digestion with ApaI. Reference supercoiled (SC) and linearized (Lin) pMP179(115 repeats) and a molecular weight DNA ladder (M) are also shown. C) Graph represents the percentage of BQQ-OP mediated cleavage obtained for pMP179(115 repeats) in the absence (indicate in the graph as pMP179) or in the presence of PNA- or DNA oligomers. Values indicate the ratio between the intensity of DNA double strand cleavage to the total band intensity from the respective lane and are expressed as mean±S.D. (n=3). No cleavage was obtained in the presence of GAA-PNA and therefore not included in the graph. ** P ≤0.01 in relation to plasmid in the absence of oligonucleotide (pMP179).

    Techniques Used: Generated, Incubation, Molecular Weight, Plasmid Preparation

    Structural- and chemical probing of DNA and DNA-PNA complex formation at (GAA) 9 repeats. Non-denaturing PAGE of DNA fragments mapped by BQQ-OP cleavage and chloroacetaldehyde (CAA) modification followed by primer extension (PE). Plasmid pMP141(9 repeats), was incubated in the absence (No PNA) or presence of 10 μM of GAA-PNA or CTT-PNA in buffer (10 mM sodium cacodylate, 100 mM NaCl, 2 mM MgCl 2 , pH 7.5). The plasmid was then either chemically modified using 2% CAA or cleaved using 1 μM BQQ-OP. Untreated plasmid was used in a set of four different controls (C) of the PE reactions: C1= plasmid incubated in the absence of PNA. C2= plasmid incubated in the presence of CTT-PNA. C3= plasmid incubated in the presence of GAA-PNA. C4= plasmid not incubated. All samples were linearized using ApaI and then used as templates for the PE reaction. Sequence ladders using dideoxynucleotides (C=ddCTP, T=ddTTP, G=ddGTP and A=ddATP), linearized pMP141(9 repeats) and a PE reaction control (C4) are also shown. The left gel panel shows the DNA fragments obtained in a PE reaction using the GAA containing (R)-strand as template, and the right gel panel shows the DNA fragments obtained when using the CTT containing (Y)-strand as template. The A 1 to A 23 nucleotides flanking the repeats of the R-strand and T 1 to T 23 flanking in the Y-strand and the mid-point of the repeat sequence (M) are indicated.
    Figure Legend Snippet: Structural- and chemical probing of DNA and DNA-PNA complex formation at (GAA) 9 repeats. Non-denaturing PAGE of DNA fragments mapped by BQQ-OP cleavage and chloroacetaldehyde (CAA) modification followed by primer extension (PE). Plasmid pMP141(9 repeats), was incubated in the absence (No PNA) or presence of 10 μM of GAA-PNA or CTT-PNA in buffer (10 mM sodium cacodylate, 100 mM NaCl, 2 mM MgCl 2 , pH 7.5). The plasmid was then either chemically modified using 2% CAA or cleaved using 1 μM BQQ-OP. Untreated plasmid was used in a set of four different controls (C) of the PE reactions: C1= plasmid incubated in the absence of PNA. C2= plasmid incubated in the presence of CTT-PNA. C3= plasmid incubated in the presence of GAA-PNA. C4= plasmid not incubated. All samples were linearized using ApaI and then used as templates for the PE reaction. Sequence ladders using dideoxynucleotides (C=ddCTP, T=ddTTP, G=ddGTP and A=ddATP), linearized pMP141(9 repeats) and a PE reaction control (C4) are also shown. The left gel panel shows the DNA fragments obtained in a PE reaction using the GAA containing (R)-strand as template, and the right gel panel shows the DNA fragments obtained when using the CTT containing (Y)-strand as template. The A 1 to A 23 nucleotides flanking the repeats of the R-strand and T 1 to T 23 flanking in the Y-strand and the mid-point of the repeat sequence (M) are indicated.

    Techniques Used: Polyacrylamide Gel Electrophoresis, Cellular Antioxidant Activity Assay, Modification, Plasmid Preparation, Incubation, Sequencing

    BQQ-OP mediated DNA cleavage of H-DNA forming (GAA) 115 repeats in the presence of LNA-PS. A) Representative gel from pMP179(115 repeats) incubated with 10 μM LNA-PS (GAA-LNA-PS 1, 2 or 3 or ER-LNA-PS or CTT-LNA-PS, lane 1, 2, 3, 4 or 7, respectively), in the absence of oligonucleotide (lane 8) or with 10 μM ssDNA (CTT-DNA or GAA-DNA, lane 5 or 6, respectively) in buffer (10 mM sodium cacodylate, 100 mM NaCl, 2 mM MgCl 2 , pH 7.5). Triplex-specific cleavage was carried out in the presence of Cu 2+ and MPA, and the plasmid was subsequently digested using ApaI. The two obtained DNA fragments have approximately 3814 and 3178 bp. Reference supercoiled (SC) and linearized (Lin) pMP179(115 repeats) and a molecular weight DNA ladder (M) are also shown. B) Graph represents the percentage of BQQ-OP mediated cleavage for pMP179(115 repeats) in the absence (indicate in the graph as pMP179) or in the presence of LNA- or DNA oligomers. Values indicate the ratio between the intensity of DNA double strand cleavage to the total band intensity from the respective lane and are expressed as mean±S.D. (n=3). *** P ≤0.001, **** P
    Figure Legend Snippet: BQQ-OP mediated DNA cleavage of H-DNA forming (GAA) 115 repeats in the presence of LNA-PS. A) Representative gel from pMP179(115 repeats) incubated with 10 μM LNA-PS (GAA-LNA-PS 1, 2 or 3 or ER-LNA-PS or CTT-LNA-PS, lane 1, 2, 3, 4 or 7, respectively), in the absence of oligonucleotide (lane 8) or with 10 μM ssDNA (CTT-DNA or GAA-DNA, lane 5 or 6, respectively) in buffer (10 mM sodium cacodylate, 100 mM NaCl, 2 mM MgCl 2 , pH 7.5). Triplex-specific cleavage was carried out in the presence of Cu 2+ and MPA, and the plasmid was subsequently digested using ApaI. The two obtained DNA fragments have approximately 3814 and 3178 bp. Reference supercoiled (SC) and linearized (Lin) pMP179(115 repeats) and a molecular weight DNA ladder (M) are also shown. B) Graph represents the percentage of BQQ-OP mediated cleavage for pMP179(115 repeats) in the absence (indicate in the graph as pMP179) or in the presence of LNA- or DNA oligomers. Values indicate the ratio between the intensity of DNA double strand cleavage to the total band intensity from the respective lane and are expressed as mean±S.D. (n=3). *** P ≤0.001, **** P

    Techniques Used: Incubation, Plasmid Preparation, Molecular Weight

    2) Product Images from "Characterization of Plasmids in Extensively Drug-Resistant Acinetobacter Strains Isolated in India and Pakistan"

    Article Title: Characterization of Plasmids in Extensively Drug-Resistant Acinetobacter Strains Isolated in India and Pakistan

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.03242-14

    (a) Pulsed-field gel of S1 nuclease-digested genomic DNA from A. bereziniae CHI-40-1, recipients, and transconjugants; (b) in-gel hybridization with a bla NDM-1 gene probe; (c) pulsed-field gel of ApaI-digested genomic DNA from CHI-40-1; (d) in-gel hybridization
    Figure Legend Snippet: (a) Pulsed-field gel of S1 nuclease-digested genomic DNA from A. bereziniae CHI-40-1, recipients, and transconjugants; (b) in-gel hybridization with a bla NDM-1 gene probe; (c) pulsed-field gel of ApaI-digested genomic DNA from CHI-40-1; (d) in-gel hybridization

    Techniques Used: Pulsed-Field Gel, Hybridization

    3) Product Images from "The CpG-sites of the CBX3 ubiquitous chromatin opening element are critical structural determinants for the anti-silencing function"

    Article Title: The CpG-sites of the CBX3 ubiquitous chromatin opening element are critical structural determinants for the anti-silencing function

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-04212-8

    CBX3 subfragments and lentiviral vectors. ( A ) Schematic representation of the CBX3 element with its CpG-sites (black bars) and deleted splice sites (red bars) as well as the CBX3 subfragments obtained by PCR (CBX3(1-339) - CBX3(340-508)) and the CBX3(503-679) subfragment generated by restriction digestion with XhoI and ApaI. ( B ) Third-generation self-inactivating (SIN) lentiviral constructs expressing an eGFP cDNA from the spleen focus forming virus (SFFV) promoter in the absence or presence of either the 1.5 kb A2UCOE, the 679 bp CBX3 or any of the CBX3 subfragments. Abbreviations: ΔLTR: Long terminal repeat harboring the SIN mutation in the U3 region of the LTR; ψ: extended encapsidation signal; RRE: Rev-response element; cPPT: central polypurine tract; GFP: (enhanced) green fluorescent protein; wPRE: woodchuck hepatitis virus post-transcriptional element.
    Figure Legend Snippet: CBX3 subfragments and lentiviral vectors. ( A ) Schematic representation of the CBX3 element with its CpG-sites (black bars) and deleted splice sites (red bars) as well as the CBX3 subfragments obtained by PCR (CBX3(1-339) - CBX3(340-508)) and the CBX3(503-679) subfragment generated by restriction digestion with XhoI and ApaI. ( B ) Third-generation self-inactivating (SIN) lentiviral constructs expressing an eGFP cDNA from the spleen focus forming virus (SFFV) promoter in the absence or presence of either the 1.5 kb A2UCOE, the 679 bp CBX3 or any of the CBX3 subfragments. Abbreviations: ΔLTR: Long terminal repeat harboring the SIN mutation in the U3 region of the LTR; ψ: extended encapsidation signal; RRE: Rev-response element; cPPT: central polypurine tract; GFP: (enhanced) green fluorescent protein; wPRE: woodchuck hepatitis virus post-transcriptional element.

    Techniques Used: Polymerase Chain Reaction, Generated, Construct, Expressing, Mutagenesis

    4) Product Images from "Targeting Activation Induced Cytidine Deaminase Overcome Tumor Evasion of Immunotherapy by Cytotoxic T Lymphocytes"

    Article Title: Targeting Activation Induced Cytidine Deaminase Overcome Tumor Evasion of Immunotherapy by Cytotoxic T Lymphocytes

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi: 10.4049/jimmunol.0903322

    Generation and characterization of AID-silenced J558 cells (AID 2.13, AID 2.36, AID 2.24, AID 2.17 and AID 2.27) and controls (C4 and C5). ApaI and EcoRI restriction sites were added to the AID targeting sequence (5'-AATTGTTGTTCCTACGCTACA-3') at 5'- and
    Figure Legend Snippet: Generation and characterization of AID-silenced J558 cells (AID 2.13, AID 2.36, AID 2.24, AID 2.17 and AID 2.27) and controls (C4 and C5). ApaI and EcoRI restriction sites were added to the AID targeting sequence (5'-AATTGTTGTTCCTACGCTACA-3') at 5'- and

    Techniques Used: Sequencing

    Related Articles

    Clone Assay:

    Article Title: Transposon mutagenesis identifies genes essential for Plasmodium falciparum gametocytogenesis
    Article Snippet: .. After the correct sequence was verified (Macrogen), the insert was excised with either KpnI or ApaI (FastDigest; Fermentas) and cloned into pINTp3 , or the insert was excised with SpeI and NotI (FastDigest; Fermentas) and cloned into pCBM.NEO , both of which confer resistance to Geneticin/G418. .. Parasites were transfected with 100 μg rescue construct or 100 μg empty vector with the drug resistance marker but no rescue insert (pINT or pCBM.NEO) as described above; 48 h posttransfection, 800–1,000 μg/mL G418 (Sigma) were added to culture, and medium was changed daily until parasites disappeared from culture.

    Agarose Gel Electrophoresis:

    Article Title: Analysis of siRNA specificity on targets with double-nucleotide mismatches
    Article Snippet: .. The purified PCR product was cleaved with restriction enzymes FastDigest ApaI and BglII (Fermentas GmbH, St. Leon-Rot, Germany) followed by analysis on a 2% MetaPhor agarose gel for verification of correct size (33 bp). .. The forward and reverse strands of the wild type target of siCD46 were ordered separately and annealed to form a ready-to-ligate target sequence.

    Isolation:

    Article Title: Disruption of Higher Order DNA Structures in Friedreich’s Ataxia (GAA)n Repeats by PNA or LNA Targeting
    Article Snippet: .. The isolated DNA was digested using ApaI (Fermentas Fastdigest) and the enzyme was then inactivated, 5 min at 65°C. .. CAA chemical modification : CAA (2%) was added to the plasmid solution (final volume 20 μl) and the reaction was allowed to proceed during 30 min at 37°C, followed by isolation of the DNA using miniprep columns (Qiagen).

    Purification:

    Article Title: Analysis of siRNA specificity on targets with double-nucleotide mismatches
    Article Snippet: .. The purified PCR product was cleaved with restriction enzymes FastDigest ApaI and BglII (Fermentas GmbH, St. Leon-Rot, Germany) followed by analysis on a 2% MetaPhor agarose gel for verification of correct size (33 bp). .. The forward and reverse strands of the wild type target of siCD46 were ordered separately and annealed to form a ready-to-ligate target sequence.

    Polymerase Chain Reaction:

    Article Title: Analysis of siRNA specificity on targets with double-nucleotide mismatches
    Article Snippet: .. The purified PCR product was cleaved with restriction enzymes FastDigest ApaI and BglII (Fermentas GmbH, St. Leon-Rot, Germany) followed by analysis on a 2% MetaPhor agarose gel for verification of correct size (33 bp). .. The forward and reverse strands of the wild type target of siCD46 were ordered separately and annealed to form a ready-to-ligate target sequence.

    Sequencing:

    Article Title: Transposon mutagenesis identifies genes essential for Plasmodium falciparum gametocytogenesis
    Article Snippet: .. After the correct sequence was verified (Macrogen), the insert was excised with either KpnI or ApaI (FastDigest; Fermentas) and cloned into pINTp3 , or the insert was excised with SpeI and NotI (FastDigest; Fermentas) and cloned into pCBM.NEO , both of which confer resistance to Geneticin/G418. .. Parasites were transfected with 100 μg rescue construct or 100 μg empty vector with the drug resistance marker but no rescue insert (pINT or pCBM.NEO) as described above; 48 h posttransfection, 800–1,000 μg/mL G418 (Sigma) were added to culture, and medium was changed daily until parasites disappeared from culture.

    Transformation Assay:

    Article Title: RifZ (AMED_0655) Is a Pathway-Specific Regulator for Rifamycin Biosynthesis in Amycolatopsis mediterranei
    Article Snippet: .. Genomic DNA was extracted using commercial genomic extraction kits and digested with FastDigest ApaI (Thermo Fisher Scientific) for about 3 h. After inactivation of ApaI, the digested mixture was ligated with T4 DNA ligase (Tolo Biotech, Shanghai, China) overnight before being transformed into competent cells of E. coli DH5α. .. Transformants selected on LB plates containing apramycin (50 μg/ml) were inoculated into 3 ml liquid LB medium and incubated overnight.

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    Thermo Fisher apai
    (a) Pulsed-field gel of <t>S1</t> nuclease-digested genomic DNA from A. bereziniae CHI-40-1, recipients, and transconjugants; (b) in-gel hybridization with a bla NDM-1 gene probe; (c) pulsed-field gel of <t>ApaI-digested</t> genomic DNA from CHI-40-1; (d) in-gel hybridization
    Apai, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 48 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/apai/product/Thermo Fisher
    Average 95 stars, based on 48 article reviews
    Price from $9.99 to $1999.99
    apai - by Bioz Stars, 2020-05
    95/100 stars
      Buy from Supplier

    99
    Thermo Fisher noti apai restriction enzymes
    (a) Pulsed-field gel of <t>S1</t> nuclease-digested genomic DNA from A. bereziniae CHI-40-1, recipients, and transconjugants; (b) in-gel hybridization with a bla NDM-1 gene probe; (c) pulsed-field gel of <t>ApaI-digested</t> genomic DNA from CHI-40-1; (d) in-gel hybridization
    Noti Apai Restriction Enzymes, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/noti apai restriction enzymes/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    noti apai restriction enzymes - by Bioz Stars, 2020-05
    99/100 stars
      Buy from Supplier

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    (a) Pulsed-field gel of S1 nuclease-digested genomic DNA from A. bereziniae CHI-40-1, recipients, and transconjugants; (b) in-gel hybridization with a bla NDM-1 gene probe; (c) pulsed-field gel of ApaI-digested genomic DNA from CHI-40-1; (d) in-gel hybridization

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Characterization of Plasmids in Extensively Drug-Resistant Acinetobacter Strains Isolated in India and Pakistan

    doi: 10.1128/AAC.03242-14

    Figure Lengend Snippet: (a) Pulsed-field gel of S1 nuclease-digested genomic DNA from A. bereziniae CHI-40-1, recipients, and transconjugants; (b) in-gel hybridization with a bla NDM-1 gene probe; (c) pulsed-field gel of ApaI-digested genomic DNA from CHI-40-1; (d) in-gel hybridization

    Article Snippet: Plugs were treated with ApaI (Thermo Scientific, Waltham, MA, USA) or S1 nuclease (Thermo Scientific) and pulsed-field gel electrophoresis (PFGE) performed as described previously ( , ).

    Techniques: Pulsed-Field Gel, Hybridization

    CBX3 subfragments and lentiviral vectors. ( A ) Schematic representation of the CBX3 element with its CpG-sites (black bars) and deleted splice sites (red bars) as well as the CBX3 subfragments obtained by PCR (CBX3(1-339) - CBX3(340-508)) and the CBX3(503-679) subfragment generated by restriction digestion with XhoI and ApaI. ( B ) Third-generation self-inactivating (SIN) lentiviral constructs expressing an eGFP cDNA from the spleen focus forming virus (SFFV) promoter in the absence or presence of either the 1.5 kb A2UCOE, the 679 bp CBX3 or any of the CBX3 subfragments. Abbreviations: ΔLTR: Long terminal repeat harboring the SIN mutation in the U3 region of the LTR; ψ: extended encapsidation signal; RRE: Rev-response element; cPPT: central polypurine tract; GFP: (enhanced) green fluorescent protein; wPRE: woodchuck hepatitis virus post-transcriptional element.

    Journal: Scientific Reports

    Article Title: The CpG-sites of the CBX3 ubiquitous chromatin opening element are critical structural determinants for the anti-silencing function

    doi: 10.1038/s41598-017-04212-8

    Figure Lengend Snippet: CBX3 subfragments and lentiviral vectors. ( A ) Schematic representation of the CBX3 element with its CpG-sites (black bars) and deleted splice sites (red bars) as well as the CBX3 subfragments obtained by PCR (CBX3(1-339) - CBX3(340-508)) and the CBX3(503-679) subfragment generated by restriction digestion with XhoI and ApaI. ( B ) Third-generation self-inactivating (SIN) lentiviral constructs expressing an eGFP cDNA from the spleen focus forming virus (SFFV) promoter in the absence or presence of either the 1.5 kb A2UCOE, the 679 bp CBX3 or any of the CBX3 subfragments. Abbreviations: ΔLTR: Long terminal repeat harboring the SIN mutation in the U3 region of the LTR; ψ: extended encapsidation signal; RRE: Rev-response element; cPPT: central polypurine tract; GFP: (enhanced) green fluorescent protein; wPRE: woodchuck hepatitis virus post-transcriptional element.

    Article Snippet: The CBX3(503-679) subfragment was generated by digesting the CBX3.SFFV.eGFP with XhoI and ApaI, filling the ends by using Klenow fragment (ThermoFisher Scientific) and subsequently self-ligating the vector to generate the construct CBX3(503-679).SFFV.eGFP.

    Techniques: Polymerase Chain Reaction, Generated, Construct, Expressing, Mutagenesis