shrimp alkaline phosphatase  (Thermo Fisher)


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    Structured Review

    Thermo Fisher shrimp alkaline phosphatase
    Shrimp Alkaline Phosphatase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 385 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/shrimp alkaline phosphatase/product/Thermo Fisher
    Average 99 stars, based on 385 article reviews
    Price from $9.99 to $1999.99
    shrimp alkaline phosphatase - by Bioz Stars, 2020-01
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Severe Combined Adrenal and Gonadal Deficiency Caused by Novel Mutations in the Cholesterol Side Chain Cleavage Enzyme, P450scc
    Article Snippet: PCR products were purified using 1 U/μl Exonuclease I (New England Biolabs, Beverly, MA) and 0.1 U/μl shrimp alkaline phosphatase (USB Corp., Cleveland, OH) and then subjected to direct sequencing using BigDye terminator v1.1 (Applied Biosystems, Foster City, CA) and a MegaBACE1000 capillary DNA sequencer (Amersham Biosciences Inc., Little Chalfont, UK). .. Compound heterozygous mutations were confirmed by studying parental DNA, as well as by subcloning PCR products into a TA-cloning vector (Invitrogen, Carlsbad, CA) and sequencing independent clones.

    Amplification:

    Article Title: A RAPGEF6 variant constitutes a major risk factor for laryngeal paralysis in dogs
    Article Snippet: A 265 bp or 301 bp fragment containing the variable position was PCR amplified (35 cycles) from genomic DNA using AmpliTaq Gold 360 Master Mix (ThermoFisher). .. After treatment with shrimp alkaline phosphatase and endonuclease I, PCR products were directly sequenced on an ABI 3730 capillary sequencer (ThermoFisher).

    Article Title: Mutation analysis of PAH gene and characterization of a recurrent deletion mutation in Korean patients with phenylketonuria
    Article Snippet: .. Five microliters of amplification product were treated with 10 U shrimp alkaline phosphatase and 2 U exonuclease I (USB Corp., Cleveland, OH), and direct sequencing was then performed using the BigDye Terminator Cycle Sequencing Ready Reaction kit (Applied Biosystems) in the ABI Prism 3100 genetic analyzer (Applied Biosystems). ..

    Article Title: Development of a Respiratory Virus Panel Test for Detection of Twenty Human Respiratory Viruses by Use of Multiplex PCR and a Fluid Microbead-Based Assay ▿
    Article Snippet: The primers were chosen carefully for target specificity and size so that amplicons would be small (100 to 400 bp) to maximize the amplification efficiency; the median amplicon size was 204 bp. .. Following PCR, the remaining dNTPs and primers were removed by incubating the entire 25-μl PCR mixture with 2.5 U shrimp alkaline phosphatase and 10 U exonuclease (Invitrogen) for 30 min at 37°C, followed by 30 s at 99°C.

    Article Title: Dysferlin, Annexin A1 and Mitsugumin-53 Are Upregulated in Muscular Dystrophy and Localize to Longitudinal Tubules of the T-System with Stretch
    Article Snippet: The amplicon for region 2 was generated using Platinum Pfx Enzyme (Invitrogen) and the amplicons for regions 3–7b were generated using Taq DNA Polymerase (Invitrogen). .. The PCR product for region 2 was gel-purified (Jetquick gel extraction spin kit, Astral Scientific, Caringbah, Australia) while products for regions 3–7b were treated with 1U Shrimp Alkaline Phosphatase (USB Corporation, Cleveland, OH) and 6U Exonuclease I (New England Biolabs, Ipswich, MA).

    Article Title: Specific and Sensitive Detection of Neisseria gonorrhoeae in Clinical Specimens by Real-Time PCR
    Article Snippet: M13- opa- Fw (TGT AAA ACG ACG GCC AGT GTT GAA ACA CCG CCC GG) and M13- opa- Rv (CAG GAA ACA GCT ATG ACC CGG TTT GAC CGG TTA AAA AAA GAT) primers (300 nM each) were used for amplification. .. The PCR product was purified by adding 4 μl of combined exonuclease I (10 U/ml) and shrimp alkaline phosphatase (2 U/μl) (USB Corporation; distributor, Amersham Bioscience, Roosendaal, The Netherlands) to 20 μl of PCR product and incubating the mixture for 15 min at 37°C, followed by inactivation for 15 min at 80°C (PTC-200 thermocycler; MJ Research, Biozym TC BV, Landgraaf, The Netherlands).

    Article Title: Autosomal dominant nanophthalmos and high hyperopia associated with a C-terminal frameshift variant in MYRF
    Article Snippet: .. The following standard PCR conditions were used on a Veriti (Thermo Fisher Scientific) thermal cycler: Step 1, 95 °C for 10 min; Step 2, 95 °C for 30 s, 55 °C for 30 s, 72 °C for 1 min, repeated for 40 cycles; and Step 3, 72 °C for 7 min. PCR amplified products were prepared for sequencing with the ExoSAP method using 10 μl of each PCR reaction treated with 5 U of Exonuclease I (New England Biolabs, Ipswich, MA) and 1 U of Shrimp Alkaline Phosphatase (USB Corporation, Cleveland, OH) to remove residual primers and dNTPs. .. Bidirectional BigDye Terminator Cycle Sequencing (Thermo Fisher Scientific) reactions of the appropriate template and PCR primer were resolved and base called on an Applied Biosystems 3130xl Genetic Analyzer (Thermo Fisher Scientific).

    Article Title: Matrix Assisted Laser Desorption Ionization-Time-of-Flight mass spectrometry identification of Mycobacterium bovis in Bovinae
    Article Snippet: For characterization of the Mb400 negative isolates, the partial sequence of the hsp65 gene ofMycobacterium sp. was amplified [ ]. .. In order to remove excess primers and unincorporated nucleotides, the products of the hsp65 PCRs were purified with the enzymes exonuclease I and shrimp alkaline phosphatase [ ], and sequenced with the BigDye Terminator cycle sequencing kit version 3.1 (Applied Biosystems, Foster City, CA, U.S.A.).

    Synthesized:

    Article Title: Development of a Respiratory Virus Panel Test for Detection of Twenty Human Respiratory Viruses by Use of Multiplex PCR and a Fluid Microbead-Based Assay ▿
    Article Snippet: For the RVP assay, a two-step reverse transcriptase PCR (RT-PCR) was used. cDNA was synthesized using Moloney murine leukemia virus RT (Invitrogen) in a 20-μl reaction mixture containing 0.5 μM random hexamers, 0.5 μM deoxynucleoside triphosphates (dNTPs), 1× RT buffer, 0.01 M dithiothreitol, and 5 μl nucleic acid for 60 min at 37°C. cDNA (5 μl) was amplified in a multiplex PCR using 14 primer pairs designed to amplify highly conserved regions of individual viral genomes for detection of specific respiratory virus types and subtypes. .. Following PCR, the remaining dNTPs and primers were removed by incubating the entire 25-μl PCR mixture with 2.5 U shrimp alkaline phosphatase and 10 U exonuclease (Invitrogen) for 30 min at 37°C, followed by 30 s at 99°C.

    TA Cloning:

    Article Title: Severe Combined Adrenal and Gonadal Deficiency Caused by Novel Mutations in the Cholesterol Side Chain Cleavage Enzyme, P450scc
    Article Snippet: PCR products were purified using 1 U/μl Exonuclease I (New England Biolabs, Beverly, MA) and 0.1 U/μl shrimp alkaline phosphatase (USB Corp., Cleveland, OH) and then subjected to direct sequencing using BigDye terminator v1.1 (Applied Biosystems, Foster City, CA) and a MegaBACE1000 capillary DNA sequencer (Amersham Biosciences Inc., Little Chalfont, UK). .. Compound heterozygous mutations were confirmed by studying parental DNA, as well as by subcloning PCR products into a TA-cloning vector (Invitrogen, Carlsbad, CA) and sequencing independent clones.

    Incubation:

    Article Title: Lethal Mutagenesis of Poliovirus Mediated by a Mutagenic Pyrimidine Analogue ▿
    Article Snippet: .. A 1/10 volume of 1 M ammonium bicarbonate (pH 7.8) was then added to the digest, followed by 1 U shrimp alkaline phosphatase (USB Corporation) and further incubation for 1 h at 37°C. .. The solution was heated to 95°C for 10 min to inactivate enzymes prior to high-performance liquid chromatography (HPLC) separation.

    Activity Assay:

    Article Title: Inhibition of the Protein Phosphatase CppA Alters Development of Chlamydia trachomatis
    Article Snippet: MDSA and various MDSA derivatives were screened against rCppA and shrimp alkaline phosphatase (SAP; Thermo Fisher). .. To test the inhibitory activity of each compound against SAP, pNPP assays were performed in the presence of either 5 mM MnCl2 or 5 mM MgCl2 .

    Mass Spectrometry:

    Article Title: The Thiamine Kinase (YcfN) Enzyme Plays a Minor but Significant Role in Cobinamide Salvaging in Salmonella enterica ▿
    Article Snippet: Paragraph title: (vi) MS. ... The remainder of the mixture was treated with shrimp alkaline phosphatase (SAP; Fermentas) according to the manufacturer's instructions, loaded onto a preequilibrated Sep-Pak C18 cartridge (Waters), and eluted and dried as described above.

    Modification:

    Article Title: Dysferlin, Annexin A1 and Mitsugumin-53 Are Upregulated in Muscular Dystrophy and Localize to Longitudinal Tubules of the T-System with Stretch
    Article Snippet: Genomic DNA was isolated from either frozen muscle using the QIAamp DNA Mini Kit (Qiagen Pty Limited, Doncaster, VIC, Australia), or from whole blood using a modified salting-out protocol ( ). .. The PCR product for region 2 was gel-purified (Jetquick gel extraction spin kit, Astral Scientific, Caringbah, Australia) while products for regions 3–7b were treated with 1U Shrimp Alkaline Phosphatase (USB Corporation, Cleveland, OH) and 6U Exonuclease I (New England Biolabs, Ipswich, MA).

    Countercurrent Chromatography:

    Article Title: Specific and Sensitive Detection of Neisseria gonorrhoeae in Clinical Specimens by Real-Time PCR
    Article Snippet: M13- opa- Fw (TGT AAA ACG ACG GCC AGT GTT GAA ACA CCG CCC GG) and M13- opa- Rv (CAG GAA ACA GCT ATG ACC CGG TTT GAC CGG TTA AAA AAA GAT) primers (300 nM each) were used for amplification. .. The PCR product was purified by adding 4 μl of combined exonuclease I (10 U/ml) and shrimp alkaline phosphatase (2 U/μl) (USB Corporation; distributor, Amersham Bioscience, Roosendaal, The Netherlands) to 20 μl of PCR product and incubating the mixture for 15 min at 37°C, followed by inactivation for 15 min at 80°C (PTC-200 thermocycler; MJ Research, Biozym TC BV, Landgraaf, The Netherlands).

    Generated:

    Article Title: Dysferlin, Annexin A1 and Mitsugumin-53 Are Upregulated in Muscular Dystrophy and Localize to Longitudinal Tubules of the T-System with Stretch
    Article Snippet: The amplicon for region 2 was generated using Platinum Pfx Enzyme (Invitrogen) and the amplicons for regions 3–7b were generated using Taq DNA Polymerase (Invitrogen). .. The PCR product for region 2 was gel-purified (Jetquick gel extraction spin kit, Astral Scientific, Caringbah, Australia) while products for regions 3–7b were treated with 1U Shrimp Alkaline Phosphatase (USB Corporation, Cleveland, OH) and 6U Exonuclease I (New England Biolabs, Ipswich, MA).

    Polymerase Chain Reaction:

    Article Title: A RAPGEF6 variant constitutes a major risk factor for laryngeal paralysis in dogs
    Article Snippet: .. After treatment with shrimp alkaline phosphatase and endonuclease I, PCR products were directly sequenced on an ABI 3730 capillary sequencer (ThermoFisher). .. We analyzed the Sanger sequence data using the software Sequencher 5.1 (GeneCodes).

    Article Title: Mutation analysis of PAH gene and characterization of a recurrent deletion mutation in Korean patients with phenylketonuria
    Article Snippet: All 13 PAH exons and their flanking intronic sequences were amplified by PCR using appropriate primers designed by the authors (sequences available upon request) and a thermal cycler (Model 9700; Applied Biosystems, Foster City, CA). .. Five microliters of amplification product were treated with 10 U shrimp alkaline phosphatase and 2 U exonuclease I (USB Corp., Cleveland, OH), and direct sequencing was then performed using the BigDye Terminator Cycle Sequencing Ready Reaction kit (Applied Biosystems) in the ABI Prism 3100 genetic analyzer (Applied Biosystems).

    Article Title: Development of a Respiratory Virus Panel Test for Detection of Twenty Human Respiratory Viruses by Use of Multiplex PCR and a Fluid Microbead-Based Assay ▿
    Article Snippet: .. Following PCR, the remaining dNTPs and primers were removed by incubating the entire 25-μl PCR mixture with 2.5 U shrimp alkaline phosphatase and 10 U exonuclease (Invitrogen) for 30 min at 37°C, followed by 30 s at 99°C. .. A multiplex target-specific primer extension (TSPE) reaction was used to detect specific viral sequences amplified by RT-PCR.

    Article Title: Dysferlin, Annexin A1 and Mitsugumin-53 Are Upregulated in Muscular Dystrophy and Localize to Longitudinal Tubules of the T-System with Stretch
    Article Snippet: .. The PCR product for region 2 was gel-purified (Jetquick gel extraction spin kit, Astral Scientific, Caringbah, Australia) while products for regions 3–7b were treated with 1U Shrimp Alkaline Phosphatase (USB Corporation, Cleveland, OH) and 6U Exonuclease I (New England Biolabs, Ipswich, MA). .. All DNA sequencing was performed on an Applied Biosystems 3730 xl capillary sequencer.

    Article Title: Severe Combined Adrenal and Gonadal Deficiency Caused by Novel Mutations in the Cholesterol Side Chain Cleavage Enzyme, P450scc
    Article Snippet: .. PCR products were purified using 1 U/μl Exonuclease I (New England Biolabs, Beverly, MA) and 0.1 U/μl shrimp alkaline phosphatase (USB Corp., Cleveland, OH) and then subjected to direct sequencing using BigDye terminator v1.1 (Applied Biosystems, Foster City, CA) and a MegaBACE1000 capillary DNA sequencer (Amersham Biosciences Inc., Little Chalfont, UK). .. Sequence Analyser v3.0 (Amersham Biosciences) and Sequencher v4.1 (Gene Codes Corp., Ann Arbor, MI) were used to analyze the data.

    Article Title: Chymotrypsin C (CTRC) alterations that diminish activity or secretion are associated with chronic pancreatitis
    Article Snippet: .. Briefly, we digested the PCR products with shrimp alkaline phosphatase and exonuclease I and performed cycle sequencing using BigDye terminator mix (Applied Biosystems). .. The reaction products were purified with ethanol precipitation or Sephadex G-50 and loaded onto an ABI 3100-Avant or an ABI 3730 sequencer (Applied Biosystems).

    Article Title: Specific and Sensitive Detection of Neisseria gonorrhoeae in Clinical Specimens by Real-Time PCR
    Article Snippet: .. The PCR product was purified by adding 4 μl of combined exonuclease I (10 U/ml) and shrimp alkaline phosphatase (2 U/μl) (USB Corporation; distributor, Amersham Bioscience, Roosendaal, The Netherlands) to 20 μl of PCR product and incubating the mixture for 15 min at 37°C, followed by inactivation for 15 min at 80°C (PTC-200 thermocycler; MJ Research, Biozym TC BV, Landgraaf, The Netherlands). ..

    Article Title: Autosomal dominant nanophthalmos and high hyperopia associated with a C-terminal frameshift variant in MYRF
    Article Snippet: .. The following standard PCR conditions were used on a Veriti (Thermo Fisher Scientific) thermal cycler: Step 1, 95 °C for 10 min; Step 2, 95 °C for 30 s, 55 °C for 30 s, 72 °C for 1 min, repeated for 40 cycles; and Step 3, 72 °C for 7 min. PCR amplified products were prepared for sequencing with the ExoSAP method using 10 μl of each PCR reaction treated with 5 U of Exonuclease I (New England Biolabs, Ipswich, MA) and 1 U of Shrimp Alkaline Phosphatase (USB Corporation, Cleveland, OH) to remove residual primers and dNTPs. .. Bidirectional BigDye Terminator Cycle Sequencing (Thermo Fisher Scientific) reactions of the appropriate template and PCR primer were resolved and base called on an Applied Biosystems 3130xl Genetic Analyzer (Thermo Fisher Scientific).

    Article Title: Matrix Assisted Laser Desorption Ionization-Time-of-Flight mass spectrometry identification of Mycobacterium bovis in Bovinae
    Article Snippet: Paragraph title: PCR, sequencing, and spoligotyping ... In order to remove excess primers and unincorporated nucleotides, the products of the hsp65 PCRs were purified with the enzymes exonuclease I and shrimp alkaline phosphatase [ ], and sequenced with the BigDye Terminator cycle sequencing kit version 3.1 (Applied Biosystems, Foster City, CA, U.S.A.).

    DNA Sequencing:

    Article Title: Dysferlin, Annexin A1 and Mitsugumin-53 Are Upregulated in Muscular Dystrophy and Localize to Longitudinal Tubules of the T-System with Stretch
    Article Snippet: The PCR product for region 2 was gel-purified (Jetquick gel extraction spin kit, Astral Scientific, Caringbah, Australia) while products for regions 3–7b were treated with 1U Shrimp Alkaline Phosphatase (USB Corporation, Cleveland, OH) and 6U Exonuclease I (New England Biolabs, Ipswich, MA). .. All DNA sequencing was performed on an Applied Biosystems 3730 xl capillary sequencer.

    Article Title: Severe Combined Adrenal and Gonadal Deficiency Caused by Novel Mutations in the Cholesterol Side Chain Cleavage Enzyme, P450scc
    Article Snippet: Mutations in StAR and SF1 were excluded by DNA sequencing, as described ( , ). .. PCR products were purified using 1 U/μl Exonuclease I (New England Biolabs, Beverly, MA) and 0.1 U/μl shrimp alkaline phosphatase (USB Corp., Cleveland, OH) and then subjected to direct sequencing using BigDye terminator v1.1 (Applied Biosystems, Foster City, CA) and a MegaBACE1000 capillary DNA sequencer (Amersham Biosciences Inc., Little Chalfont, UK).

    Article Title: Autosomal dominant nanophthalmos and high hyperopia associated with a C-terminal frameshift variant in MYRF
    Article Snippet: Paragraph title: DNA sequencing and analysis ... The following standard PCR conditions were used on a Veriti (Thermo Fisher Scientific) thermal cycler: Step 1, 95 °C for 10 min; Step 2, 95 °C for 30 s, 55 °C for 30 s, 72 °C for 1 min, repeated for 40 cycles; and Step 3, 72 °C for 7 min. PCR amplified products were prepared for sequencing with the ExoSAP method using 10 μl of each PCR reaction treated with 5 U of Exonuclease I (New England Biolabs, Ipswich, MA) and 1 U of Shrimp Alkaline Phosphatase (USB Corporation, Cleveland, OH) to remove residual primers and dNTPs.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Development of a Respiratory Virus Panel Test for Detection of Twenty Human Respiratory Viruses by Use of Multiplex PCR and a Fluid Microbead-Based Assay ▿
    Article Snippet: Paragraph title: RT-PCR. ... Following PCR, the remaining dNTPs and primers were removed by incubating the entire 25-μl PCR mixture with 2.5 U shrimp alkaline phosphatase and 10 U exonuclease (Invitrogen) for 30 min at 37°C, followed by 30 s at 99°C.

    High Performance Liquid Chromatography:

    Article Title: Lethal Mutagenesis of Poliovirus Mediated by a Mutagenic Pyrimidine Analogue ▿
    Article Snippet: A 1/10 volume of 1 M ammonium bicarbonate (pH 7.8) was then added to the digest, followed by 1 U shrimp alkaline phosphatase (USB Corporation) and further incubation for 1 h at 37°C. .. The solution was heated to 95°C for 10 min to inactivate enzymes prior to high-performance liquid chromatography (HPLC) separation.

    Nucleic Acid Electrophoresis:

    Article Title: A RAPGEF6 variant constitutes a major risk factor for laryngeal paralysis in dogs
    Article Snippet: After treatment with shrimp alkaline phosphatase and endonuclease I, PCR products were directly sequenced on an ABI 3730 capillary sequencer (ThermoFisher). .. To genotype larger numbers of samples, we performed fragment size analysis on a Fragment Analyzer capillary gel electrophoresis instrument (Advance Analytical).

    Mutagenesis:

    Article Title: Mutation analysis of PAH gene and characterization of a recurrent deletion mutation in Korean patients with phenylketonuria
    Article Snippet: Paragraph title: Mutation analysis ... Five microliters of amplification product were treated with 10 U shrimp alkaline phosphatase and 2 U exonuclease I (USB Corp., Cleveland, OH), and direct sequencing was then performed using the BigDye Terminator Cycle Sequencing Ready Reaction kit (Applied Biosystems) in the ABI Prism 3100 genetic analyzer (Applied Biosystems).

    Article Title: Severe Combined Adrenal and Gonadal Deficiency Caused by Novel Mutations in the Cholesterol Side Chain Cleavage Enzyme, P450scc
    Article Snippet: PCR products were purified using 1 U/μl Exonuclease I (New England Biolabs, Beverly, MA) and 0.1 U/μl shrimp alkaline phosphatase (USB Corp., Cleveland, OH) and then subjected to direct sequencing using BigDye terminator v1.1 (Applied Biosystems, Foster City, CA) and a MegaBACE1000 capillary DNA sequencer (Amersham Biosciences Inc., Little Chalfont, UK). .. Genscan splice prediction software ( ) was used to assess the consequences of the splice site mutation detected.

    Article Title: Chymotrypsin C (CTRC) alterations that diminish activity or secretion are associated with chronic pancreatitis
    Article Snippet: Paragraph title: Mutation screening ... Briefly, we digested the PCR products with shrimp alkaline phosphatase and exonuclease I and performed cycle sequencing using BigDye terminator mix (Applied Biosystems).

    Article Title: Autosomal dominant nanophthalmos and high hyperopia associated with a C-terminal frameshift variant in MYRF
    Article Snippet: The following standard PCR conditions were used on a Veriti (Thermo Fisher Scientific) thermal cycler: Step 1, 95 °C for 10 min; Step 2, 95 °C for 30 s, 55 °C for 30 s, 72 °C for 1 min, repeated for 40 cycles; and Step 3, 72 °C for 7 min. PCR amplified products were prepared for sequencing with the ExoSAP method using 10 μl of each PCR reaction treated with 5 U of Exonuclease I (New England Biolabs, Ipswich, MA) and 1 U of Shrimp Alkaline Phosphatase (USB Corporation, Cleveland, OH) to remove residual primers and dNTPs. .. Detection of sequence variants was performed with the aid of Mutation Surveyor v4.0 (SoftGenetics LLC, State College, PA), with trace files assembled against the MYRF ( NM_001127392.3 ) hg19 reference.

    Isolation:

    Article Title: Mutation analysis of PAH gene and characterization of a recurrent deletion mutation in Korean patients with phenylketonuria
    Article Snippet: Genomic DNA was isolated from peripheral blood leukocytes using the Wizard Genomic DNA Purification kit (Promega, Madison, WI)( ). .. Five microliters of amplification product were treated with 10 U shrimp alkaline phosphatase and 2 U exonuclease I (USB Corp., Cleveland, OH), and direct sequencing was then performed using the BigDye Terminator Cycle Sequencing Ready Reaction kit (Applied Biosystems) in the ABI Prism 3100 genetic analyzer (Applied Biosystems).

    Article Title: Dysferlin, Annexin A1 and Mitsugumin-53 Are Upregulated in Muscular Dystrophy and Localize to Longitudinal Tubules of the T-System with Stretch
    Article Snippet: Genomic DNA was isolated from either frozen muscle using the QIAamp DNA Mini Kit (Qiagen Pty Limited, Doncaster, VIC, Australia), or from whole blood using a modified salting-out protocol ( ). .. The PCR product for region 2 was gel-purified (Jetquick gel extraction spin kit, Astral Scientific, Caringbah, Australia) while products for regions 3–7b were treated with 1U Shrimp Alkaline Phosphatase (USB Corporation, Cleveland, OH) and 6U Exonuclease I (New England Biolabs, Ipswich, MA).

    Subcloning:

    Article Title: Severe Combined Adrenal and Gonadal Deficiency Caused by Novel Mutations in the Cholesterol Side Chain Cleavage Enzyme, P450scc
    Article Snippet: PCR products were purified using 1 U/μl Exonuclease I (New England Biolabs, Beverly, MA) and 0.1 U/μl shrimp alkaline phosphatase (USB Corp., Cleveland, OH) and then subjected to direct sequencing using BigDye terminator v1.1 (Applied Biosystems, Foster City, CA) and a MegaBACE1000 capillary DNA sequencer (Amersham Biosciences Inc., Little Chalfont, UK). .. Compound heterozygous mutations were confirmed by studying parental DNA, as well as by subcloning PCR products into a TA-cloning vector (Invitrogen, Carlsbad, CA) and sequencing independent clones.

    Purification:

    Article Title: Inhibition of the Protein Phosphatase CppA Alters Development of Chlamydia trachomatis
    Article Snippet: MDSA and various MDSA derivatives were screened against rCppA and shrimp alkaline phosphatase (SAP; Thermo Fisher). .. Phosphatase assays with purified rCppA were initiated by the addition of 3 μg purified protein to the assay buffer containing pNPP and MnCl2 .

    Article Title: The Thiamine Kinase (YcfN) Enzyme Plays a Minor but Significant Role in Cobinamide Salvaging in Salmonella enterica ▿
    Article Snippet: The remaining product of the in vitro Cbi kinase reaction mixture (15 μl) was loaded onto a Sep-Pak C18 cartridge (Waters) preequilibrated with ddH2 O. Purified corrinoids were eluted off the resin with 2.5 ml of 100% (vol/vol) methanol, and the sample was dried under a vacuum in a SpeedVac concentrator (Thermo Savant). .. The remainder of the mixture was treated with shrimp alkaline phosphatase (SAP; Fermentas) according to the manufacturer's instructions, loaded onto a preequilibrated Sep-Pak C18 cartridge (Waters), and eluted and dried as described above.

    Article Title: Severe Combined Adrenal and Gonadal Deficiency Caused by Novel Mutations in the Cholesterol Side Chain Cleavage Enzyme, P450scc
    Article Snippet: .. PCR products were purified using 1 U/μl Exonuclease I (New England Biolabs, Beverly, MA) and 0.1 U/μl shrimp alkaline phosphatase (USB Corp., Cleveland, OH) and then subjected to direct sequencing using BigDye terminator v1.1 (Applied Biosystems, Foster City, CA) and a MegaBACE1000 capillary DNA sequencer (Amersham Biosciences Inc., Little Chalfont, UK). .. Sequence Analyser v3.0 (Amersham Biosciences) and Sequencher v4.1 (Gene Codes Corp., Ann Arbor, MI) were used to analyze the data.

    Article Title: Chymotrypsin C (CTRC) alterations that diminish activity or secretion are associated with chronic pancreatitis
    Article Snippet: Briefly, we digested the PCR products with shrimp alkaline phosphatase and exonuclease I and performed cycle sequencing using BigDye terminator mix (Applied Biosystems). .. The reaction products were purified with ethanol precipitation or Sephadex G-50 and loaded onto an ABI 3100-Avant or an ABI 3730 sequencer (Applied Biosystems).

    Article Title: Specific and Sensitive Detection of Neisseria gonorrhoeae in Clinical Specimens by Real-Time PCR
    Article Snippet: .. The PCR product was purified by adding 4 μl of combined exonuclease I (10 U/ml) and shrimp alkaline phosphatase (2 U/μl) (USB Corporation; distributor, Amersham Bioscience, Roosendaal, The Netherlands) to 20 μl of PCR product and incubating the mixture for 15 min at 37°C, followed by inactivation for 15 min at 80°C (PTC-200 thermocycler; MJ Research, Biozym TC BV, Landgraaf, The Netherlands). ..

    Article Title: Matrix Assisted Laser Desorption Ionization-Time-of-Flight mass spectrometry identification of Mycobacterium bovis in Bovinae
    Article Snippet: .. In order to remove excess primers and unincorporated nucleotides, the products of the hsp65 PCRs were purified with the enzymes exonuclease I and shrimp alkaline phosphatase [ ], and sequenced with the BigDye Terminator cycle sequencing kit version 3.1 (Applied Biosystems, Foster City, CA, U.S.A.). .. Protein extraction and MALDI-TOF analysis In previous studies in our laboratory, a protein extraction method called MycoLyser was developed using the M. tuberculosis strain.

    Article Title: Probing the Mechanisms of DEAD-box Proteins as General RNA Chaperones: The C-terminal Domain of CYT-19 Mediates General Recognition of RNA †
    Article Snippet: .. The ribozyme was 5′-end-labeled by incubating with shrimp alkaline phosphatase (Fermentas, Hanover MD) and then with [γ-32 P]ATP and T4 polynucleotide kinase (New England Biolabs, Ipswich, MA) and purified by non-denaturing polyacrylamide gel electrophoresis (PAGE) as described ( ). .. Oligonucleotides were 5′-end-labeled with T4 polynucleotide kinase and purified by non-denaturing PAGE ( ).

    Sequencing:

    Article Title: A RAPGEF6 variant constitutes a major risk factor for laryngeal paralysis in dogs
    Article Snippet: Paragraph title: Sanger sequencing and targeted genotyping ... After treatment with shrimp alkaline phosphatase and endonuclease I, PCR products were directly sequenced on an ABI 3730 capillary sequencer (ThermoFisher).

    Article Title: Mutation analysis of PAH gene and characterization of a recurrent deletion mutation in Korean patients with phenylketonuria
    Article Snippet: .. Five microliters of amplification product were treated with 10 U shrimp alkaline phosphatase and 2 U exonuclease I (USB Corp., Cleveland, OH), and direct sequencing was then performed using the BigDye Terminator Cycle Sequencing Ready Reaction kit (Applied Biosystems) in the ABI Prism 3100 genetic analyzer (Applied Biosystems). ..

    Article Title: Dysferlin, Annexin A1 and Mitsugumin-53 Are Upregulated in Muscular Dystrophy and Localize to Longitudinal Tubules of the T-System with Stretch
    Article Snippet: Healthy controls for MG53 sequencing included 200 echo- and electrocardiographic normal samples from Olmsted County Heart Study (mostly Caucasian), and a panel of 100 African-American blood donors from the Coriell Institute Repository (Camden, NJ). .. The PCR product for region 2 was gel-purified (Jetquick gel extraction spin kit, Astral Scientific, Caringbah, Australia) while products for regions 3–7b were treated with 1U Shrimp Alkaline Phosphatase (USB Corporation, Cleveland, OH) and 6U Exonuclease I (New England Biolabs, Ipswich, MA).

    Article Title: Severe Combined Adrenal and Gonadal Deficiency Caused by Novel Mutations in the Cholesterol Side Chain Cleavage Enzyme, P450scc
    Article Snippet: .. PCR products were purified using 1 U/μl Exonuclease I (New England Biolabs, Beverly, MA) and 0.1 U/μl shrimp alkaline phosphatase (USB Corp., Cleveland, OH) and then subjected to direct sequencing using BigDye terminator v1.1 (Applied Biosystems, Foster City, CA) and a MegaBACE1000 capillary DNA sequencer (Amersham Biosciences Inc., Little Chalfont, UK). .. Sequence Analyser v3.0 (Amersham Biosciences) and Sequencher v4.1 (Gene Codes Corp., Ann Arbor, MI) were used to analyze the data.

    Article Title: Chymotrypsin C (CTRC) alterations that diminish activity or secretion are associated with chronic pancreatitis
    Article Snippet: .. Briefly, we digested the PCR products with shrimp alkaline phosphatase and exonuclease I and performed cycle sequencing using BigDye terminator mix (Applied Biosystems). .. The reaction products were purified with ethanol precipitation or Sephadex G-50 and loaded onto an ABI 3100-Avant or an ABI 3730 sequencer (Applied Biosystems).

    Article Title: Specific and Sensitive Detection of Neisseria gonorrhoeae in Clinical Specimens by Real-Time PCR
    Article Snippet: Paragraph title: Sequence analysis. ... The PCR product was purified by adding 4 μl of combined exonuclease I (10 U/ml) and shrimp alkaline phosphatase (2 U/μl) (USB Corporation; distributor, Amersham Bioscience, Roosendaal, The Netherlands) to 20 μl of PCR product and incubating the mixture for 15 min at 37°C, followed by inactivation for 15 min at 80°C (PTC-200 thermocycler; MJ Research, Biozym TC BV, Landgraaf, The Netherlands).

    Article Title: Autosomal dominant nanophthalmos and high hyperopia associated with a C-terminal frameshift variant in MYRF
    Article Snippet: .. The following standard PCR conditions were used on a Veriti (Thermo Fisher Scientific) thermal cycler: Step 1, 95 °C for 10 min; Step 2, 95 °C for 30 s, 55 °C for 30 s, 72 °C for 1 min, repeated for 40 cycles; and Step 3, 72 °C for 7 min. PCR amplified products were prepared for sequencing with the ExoSAP method using 10 μl of each PCR reaction treated with 5 U of Exonuclease I (New England Biolabs, Ipswich, MA) and 1 U of Shrimp Alkaline Phosphatase (USB Corporation, Cleveland, OH) to remove residual primers and dNTPs. .. Bidirectional BigDye Terminator Cycle Sequencing (Thermo Fisher Scientific) reactions of the appropriate template and PCR primer were resolved and base called on an Applied Biosystems 3130xl Genetic Analyzer (Thermo Fisher Scientific).

    Article Title: Matrix Assisted Laser Desorption Ionization-Time-of-Flight mass spectrometry identification of Mycobacterium bovis in Bovinae
    Article Snippet: .. In order to remove excess primers and unincorporated nucleotides, the products of the hsp65 PCRs were purified with the enzymes exonuclease I and shrimp alkaline phosphatase [ ], and sequenced with the BigDye Terminator cycle sequencing kit version 3.1 (Applied Biosystems, Foster City, CA, U.S.A.). .. Protein extraction and MALDI-TOF analysis In previous studies in our laboratory, a protein extraction method called MycoLyser was developed using the M. tuberculosis strain.

    Polyacrylamide Gel Electrophoresis:

    Article Title: Probing the Mechanisms of DEAD-box Proteins as General RNA Chaperones: The C-terminal Domain of CYT-19 Mediates General Recognition of RNA †
    Article Snippet: .. The ribozyme was 5′-end-labeled by incubating with shrimp alkaline phosphatase (Fermentas, Hanover MD) and then with [γ-32 P]ATP and T4 polynucleotide kinase (New England Biolabs, Ipswich, MA) and purified by non-denaturing polyacrylamide gel electrophoresis (PAGE) as described ( ). .. Oligonucleotides were 5′-end-labeled with T4 polynucleotide kinase and purified by non-denaturing PAGE ( ).

    Salting Out:

    Article Title: Dysferlin, Annexin A1 and Mitsugumin-53 Are Upregulated in Muscular Dystrophy and Localize to Longitudinal Tubules of the T-System with Stretch
    Article Snippet: Genomic DNA was isolated from either frozen muscle using the QIAamp DNA Mini Kit (Qiagen Pty Limited, Doncaster, VIC, Australia), or from whole blood using a modified salting-out protocol ( ). .. The PCR product for region 2 was gel-purified (Jetquick gel extraction spin kit, Astral Scientific, Caringbah, Australia) while products for regions 3–7b were treated with 1U Shrimp Alkaline Phosphatase (USB Corporation, Cleveland, OH) and 6U Exonuclease I (New England Biolabs, Ipswich, MA).

    IA:

    Article Title: Probing the Mechanisms of DEAD-box Proteins as General RNA Chaperones: The C-terminal Domain of CYT-19 Mediates General Recognition of RNA †
    Article Snippet: RNA and DNA oligonucleotides were purchased from Dharmacon (Lafayette, CO) and IDT (Coralville, IA), respectively. .. The ribozyme was 5′-end-labeled by incubating with shrimp alkaline phosphatase (Fermentas, Hanover MD) and then with [γ-32 P]ATP and T4 polynucleotide kinase (New England Biolabs, Ipswich, MA) and purified by non-denaturing polyacrylamide gel electrophoresis (PAGE) as described ( ).

    Plasmid Preparation:

    Article Title: Severe Combined Adrenal and Gonadal Deficiency Caused by Novel Mutations in the Cholesterol Side Chain Cleavage Enzyme, P450scc
    Article Snippet: PCR products were purified using 1 U/μl Exonuclease I (New England Biolabs, Beverly, MA) and 0.1 U/μl shrimp alkaline phosphatase (USB Corp., Cleveland, OH) and then subjected to direct sequencing using BigDye terminator v1.1 (Applied Biosystems, Foster City, CA) and a MegaBACE1000 capillary DNA sequencer (Amersham Biosciences Inc., Little Chalfont, UK). .. Compound heterozygous mutations were confirmed by studying parental DNA, as well as by subcloning PCR products into a TA-cloning vector (Invitrogen, Carlsbad, CA) and sequencing independent clones.

    Article Title: Probing the Mechanisms of DEAD-box Proteins as General RNA Chaperones: The C-terminal Domain of CYT-19 Mediates General Recognition of RNA †
    Article Snippet: The Tetrahymena ribozyme was transcribed from ScaI-digested plasmid using T7 RNA polymerase ( ) and purified using a Qiagen RNeasy column ( ). .. The ribozyme was 5′-end-labeled by incubating with shrimp alkaline phosphatase (Fermentas, Hanover MD) and then with [γ-32 P]ATP and T4 polynucleotide kinase (New England Biolabs, Ipswich, MA) and purified by non-denaturing polyacrylamide gel electrophoresis (PAGE) as described ( ).

    Software:

    Article Title: A RAPGEF6 variant constitutes a major risk factor for laryngeal paralysis in dogs
    Article Snippet: After treatment with shrimp alkaline phosphatase and endonuclease I, PCR products were directly sequenced on an ABI 3730 capillary sequencer (ThermoFisher). .. We analyzed the Sanger sequence data using the software Sequencher 5.1 (GeneCodes).

    Article Title: Severe Combined Adrenal and Gonadal Deficiency Caused by Novel Mutations in the Cholesterol Side Chain Cleavage Enzyme, P450scc
    Article Snippet: PCR products were purified using 1 U/μl Exonuclease I (New England Biolabs, Beverly, MA) and 0.1 U/μl shrimp alkaline phosphatase (USB Corp., Cleveland, OH) and then subjected to direct sequencing using BigDye terminator v1.1 (Applied Biosystems, Foster City, CA) and a MegaBACE1000 capillary DNA sequencer (Amersham Biosciences Inc., Little Chalfont, UK). .. Genscan splice prediction software ( ) was used to assess the consequences of the splice site mutation detected.

    Multiplex Assay:

    Article Title: Development of a Respiratory Virus Panel Test for Detection of Twenty Human Respiratory Viruses by Use of Multiplex PCR and a Fluid Microbead-Based Assay ▿
    Article Snippet: The multiplex PCR targeted the following viral genes: the matrix gene of influenza A virus; the three hemagglutinin genes of influenza A virus subtypes H1, H3, and H5; the prehemagglutinin gene of influenza B virus; the hemagglutinin gene of parainfluenza virus types 1 through 3; the phosphoprotein gene of parainfluenza virus type 4; the polymerase gene of respiratory syncytial virus (RSV) types A and B; the hexon gene of adenovirus; the 5′ untranslated region of enterovirus/rhinovirus; the nucleoprotein gene of MPV; and the polymerase gene of CoV. .. Following PCR, the remaining dNTPs and primers were removed by incubating the entire 25-μl PCR mixture with 2.5 U shrimp alkaline phosphatase and 10 U exonuclease (Invitrogen) for 30 min at 37°C, followed by 30 s at 99°C.

    Agarose Gel Electrophoresis:

    Article Title: Dysferlin, Annexin A1 and Mitsugumin-53 Are Upregulated in Muscular Dystrophy and Localize to Longitudinal Tubules of the T-System with Stretch
    Article Snippet: PCR fragments were resolved on a 1% agarose gel and visualized with ethidium bromide. .. The PCR product for region 2 was gel-purified (Jetquick gel extraction spin kit, Astral Scientific, Caringbah, Australia) while products for regions 3–7b were treated with 1U Shrimp Alkaline Phosphatase (USB Corporation, Cleveland, OH) and 6U Exonuclease I (New England Biolabs, Ipswich, MA).

    In Vitro:

    Article Title: The Thiamine Kinase (YcfN) Enzyme Plays a Minor but Significant Role in Cobinamide Salvaging in Salmonella enterica ▿
    Article Snippet: The remaining product of the in vitro Cbi kinase reaction mixture (15 μl) was loaded onto a Sep-Pak C18 cartridge (Waters) preequilibrated with ddH2 O. Purified corrinoids were eluted off the resin with 2.5 ml of 100% (vol/vol) methanol, and the sample was dried under a vacuum in a SpeedVac concentrator (Thermo Savant). .. The remainder of the mixture was treated with shrimp alkaline phosphatase (SAP; Fermentas) according to the manufacturer's instructions, loaded onto a preequilibrated Sep-Pak C18 cartridge (Waters), and eluted and dried as described above.

    Ethanol Precipitation:

    Article Title: Chymotrypsin C (CTRC) alterations that diminish activity or secretion are associated with chronic pancreatitis
    Article Snippet: Briefly, we digested the PCR products with shrimp alkaline phosphatase and exonuclease I and performed cycle sequencing using BigDye terminator mix (Applied Biosystems). .. The reaction products were purified with ethanol precipitation or Sephadex G-50 and loaded onto an ABI 3100-Avant or an ABI 3730 sequencer (Applied Biosystems).

    Concentration Assay:

    Article Title: Inhibition of the Protein Phosphatase CppA Alters Development of Chlamydia trachomatis
    Article Snippet: MDSA and various MDSA derivatives were screened against rCppA and shrimp alkaline phosphatase (SAP; Thermo Fisher). .. Each compound was dissolved in dimethyl sulfoxide (DMSO) to yield a 10 mM stock solution and tested at a final concentration of 18.2 μM.

    DNA Purification:

    Article Title: Mutation analysis of PAH gene and characterization of a recurrent deletion mutation in Korean patients with phenylketonuria
    Article Snippet: Genomic DNA was isolated from peripheral blood leukocytes using the Wizard Genomic DNA Purification kit (Promega, Madison, WI)( ). .. Five microliters of amplification product were treated with 10 U shrimp alkaline phosphatase and 2 U exonuclease I (USB Corp., Cleveland, OH), and direct sequencing was then performed using the BigDye Terminator Cycle Sequencing Ready Reaction kit (Applied Biosystems) in the ABI Prism 3100 genetic analyzer (Applied Biosystems).

    Gel Extraction:

    Article Title: Dysferlin, Annexin A1 and Mitsugumin-53 Are Upregulated in Muscular Dystrophy and Localize to Longitudinal Tubules of the T-System with Stretch
    Article Snippet: .. The PCR product for region 2 was gel-purified (Jetquick gel extraction spin kit, Astral Scientific, Caringbah, Australia) while products for regions 3–7b were treated with 1U Shrimp Alkaline Phosphatase (USB Corporation, Cleveland, OH) and 6U Exonuclease I (New England Biolabs, Ipswich, MA). .. All DNA sequencing was performed on an Applied Biosystems 3730 xl capillary sequencer.

    Variant Assay:

    Article Title: A RAPGEF6 variant constitutes a major risk factor for laryngeal paralysis in dogs
    Article Snippet: Sanger sequencing and targeted genotyping We used Sanger sequencing to confirm the RAPGEF6 :c.1793_1794ins36 candidate variant. .. After treatment with shrimp alkaline phosphatase and endonuclease I, PCR products were directly sequenced on an ABI 3730 capillary sequencer (ThermoFisher).

    Article Title: Autosomal dominant nanophthalmos and high hyperopia associated with a C-terminal frameshift variant in MYRF
    Article Snippet: The following standard PCR conditions were used on a Veriti (Thermo Fisher Scientific) thermal cycler: Step 1, 95 °C for 10 min; Step 2, 95 °C for 30 s, 55 °C for 30 s, 72 °C for 1 min, repeated for 40 cycles; and Step 3, 72 °C for 7 min. PCR amplified products were prepared for sequencing with the ExoSAP method using 10 μl of each PCR reaction treated with 5 U of Exonuclease I (New England Biolabs, Ipswich, MA) and 1 U of Shrimp Alkaline Phosphatase (USB Corporation, Cleveland, OH) to remove residual primers and dNTPs. .. The variant was deposited in ClinVar , with accession number VCV000635185.1.

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    Thermo Fisher shrimp alkaline phosphatase sap
    Shrimp Alkaline Phosphatase Sap, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 392 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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