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GE Healthcare exonuclease i
Exonuclease I, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 91/100, based on 54 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/exonuclease i/product/GE Healthcare
Average 91 stars, based on 54 article reviews
Price from $9.99 to $1999.99
exonuclease i - by Bioz Stars, 2019-12
91/100 stars

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Clone Assay:

Article Title: Endogenous TCR Recombination in TCR Tg Single RAG-Deficient Mice Uncovered by Robust In Vivo T Cell Activation and Selection
Article Snippet: PCR products were then incubated with 0.5 U of shrimp alkaline phosphatase and 5 U of exonuclease I (GE Healthcare) at 37°C for 40 min, followed by 20 min at 80°C. .. PCR products were then incubated with 0.5 U of shrimp alkaline phosphatase and 5 U of exonuclease I (GE Healthcare) at 37°C for 40 min, followed by 20 min at 80°C.

Amplification:

Article Title: The human squamous oesophagus has widespread capacity for clonal expansion from cells at diverse stages of differentiation
Article Snippet: Incubation conditions were as follows: 15 min at 50°C for RT, 2 min at 95°C for Taq inactivation, followed by a touch-down PCR amplification with 20 cycles of 15 s at 95°C for denaturation and 4 min for annealing and extension (decreasing temperature from 70°C down to 60°C over 5 cycles, followed by 15 cycles at 60°C) . .. Amplification was followed by DNA digestion with Exonuclease 1 (Exostar-1 step, GE Healthcare) as per the manufacturer's recommendation. qPCR was performed with LightCycler 480 SYBR Green I master (Roche Diagnostics Ltd, Rotkreuz, Switzerland) and 0.2 µM forward and reverse primer in a final volume of 10 µL using LightCycler 480 technology (Roche). qPCR consisted of 55 cycles of denaturation at 95°C (10 s), annealing at 60°C (15 s) and extension at 72°C (15 s). .. Primer sequences are provided in online supplementary table S2.

Article Title: DNA damage predicts prognosis and treatment response in colorectal liver metastases superior to immunogenic cell death and T cells
Article Snippet: Briefly, genomic DNA was extracted from tissue blocks and exon 2 and 3 of the KRAS gene and exon 15 of the BRAF gene were polymerase chain reaction (PCR) amplified with AmpliTaq Gold® DNA polymerase (Applied Biosystems, Fisher Scientific, Vienna, Austria) and corresponding oligonucleotide primers ( Table ). .. Excess of primers and deoxynucleotides (dNTPs) was removed by incubation of 5 µL PCR product with 2.5 U exonuclease I (GE Healthcare Life Sciences, Vienna, Austria) and 2.5 U shrimp alkaline phosphatase (SAP; GE Healthcare Life Sciences) at 37°C for 1 h. Enzymes were further heat inactivated at 70°C for 15 min, following sequence analysis with 1-2 µL of the purified PCR product and 4 pmol primers using the BigDye™ Terminator Cycle Sequencing Kit (Applied Biosystems) according to the manufacturer's instructions.

Article Title: Ancient female philopatry, asymmetric male gene flow, and synchronous population expansion support the influence of climatic oscillations on the evolution of South American sea lion (Otaria flavescens)
Article Snippet: Amplifications for CR and cytb were carried out in 20 μl with the following conditions: 1.5 mM MgCl2, 200 μM of each dNTP, 0.1 μM of each primer, 1 U of Platinum Taq DNA polymerase (Invitrogen), 1X PCR buffer (Invitrogen), 0.2%–0.4% Triton and 2 μl of DNA (approximately 50 ng). .. Amplification products were purified with shrimp alkaline phosphatase and exonuclease I (Amersham Biosciences). .. The purified products were sequenced in both directions using the DYEnamic ET Dye Terminator Cycle Sequencing Kit (Amersham Biosciences) and run in a MegaBace 1000 automated sequencer (Amersham Biosciences).

Article Title: Complete mitochondrial DNA sequence of the endangered fish (Bahabataipingensis): Mitogenome characterization and phylogenetic implications
Article Snippet: Paragraph title: Mitochondrial DNA amplification ... PCR products were cleaned by adding 0.45 µl of Shrimp Alkaline Phosphatase (Biotech Pharmacon), 0.9 µl of Exonuclease I (GE Healthcare) and 1.65 µl of sterile distilled H2 O to 9 µL of PCR product and incubating at 37 °C for 30 min and 80 °C for 20 min.

Article Title: Consuming viscous prey: a novel protein-secreting delivery system in neotropical snail-eating snakes
Article Snippet: Primers and PCR protocols for partial amplification of genes 12S, 16S, cytb, bdnf, and c-mos were those described in Grazziotin et al. [ ]. .. PCRs were purified with shrimp alkaline phosphatase and exonuclease I (GE Healthcare, Piscataway, NJ, USA) and sequences were processed using the DYEnamic ET Dye Terminator Cycle Sequencing Kit in a MegaBACE 1000 automated sequencer (GE Healthcare) following the manufacturer’s protocols.

Article Title: Behavioural and Genetic Characteristics of a New Species of Nasonia
Article Snippet: NCBI accession numbers for each fragment from all the strains are summarized in . .. To clean the reactions before sequencing, amplified reactions (8uL) were incubated with 0.5 U shrimp alkaline phosphatase and 1.0 U of exonuclease I (Amersham, Piscataway, NJ) along with the supplied buffer. .. Sequencing was performed directly from the amplified products using a BigDye v3.0 terminator sequencing kit and an ABI 3700 or 3730 xl (Applied Biosystems, Foster City, CA) automated sequencer.

Article Title: Specific variants in WDR35 cause a distinctive form of Ellis-van Creveld syndrome by disrupting the recruitment of the EvC complex and SMO into the cilium
Article Snippet: WDR 35 coding exons and at least 100 bp of the adjacent introns were amplified by standard PCR from peripheral blood genomic DNA. .. PCR products were enzymatically treated with alkaline phosphatase and exonuclease I (ExoSapit; GE Healthcare) prior to be sequenced using a dye terminator cycle sequencing kit and run on an ABI 3730 sequencer (Applied Biosystems).

Article Title: Identification of a Mutation Causing Deficient BMP1/mTLD Proteolytic Activity in Autosomal Recessive Osteogenesis Imperfecta
Article Snippet: BMP1 coding exons and at least 100 bp of the flanking introns were amplified by standard PCR (primers available on request). .. Prior to sequencing, the PCR products were treated with shrimp alkaline phosphatase and exonuclease I (ExoSapit; GE Healthcare, Piscataway, NJ) according to the manufacturers instructions.

Article Title: FGFR3 protein expression and its relationship to mutation status and prognostic variables in bladder cancer
Article Snippet: 2 μl samples of extracted DNA were amplified in 50 μl reactions using AmpliTaq Gold (Applied Biosystems, Foster City, CA, USA). .. For sequencing, unincorporated primers and deoxynucleotides were removed using shrimp alkaline phosphatase and exonuclease I (Amersham Biosciences, Chalfont, UK).

Article Title: Evolution of Smooth Tubercle Bacilli PE and PE_PGRS Genes: Evidence for a Prominent Role of Recombination and Imprint of Positive Selection
Article Snippet: Paragraph title: PCR amplification and DNA sequencing ... Amplicons were subjected to sequencing after treatment with Exonuclease I (Amersham Biosciences) and Shrimp Alkaline Phosphatase (Amersham Biosciences).

Article Title: Association of Mitochondrial DNA Polymerase ? Gene POLG1 Polymorphisms with Parkinsonism in Chinese Populations
Article Snippet: PCR conditions were set as follows: 2 min initial denaturation at 95°C, 30 cycles of 30 s denaturation at 94°C, 30 s annealing at 53–60°C, 30 s extension at 72°C and 10 min final extension at 72°C. .. Before sequencing, amplification products were incubated with shrimp alkaline phosphatase (0.5 units; Amersham) and exonuclease I (1 units; Amersham) at 37°C for 30 min, followed by heat inactivation at 80°C for 15 min. .. The DNA samples containing extension products were analyzed using the sequencing kit ABI PRISM dye terminator cycle and the automatic sequencing system ABI 3100 (Applied Biosystems).

Article Title: Endogenous TCR Recombination in TCR Tg Single RAG-Deficient Mice Uncovered by Robust In Vivo T Cell Activation and Selection
Article Snippet: PCR products were then incubated with 0.5 U of shrimp alkaline phosphatase and 5 U of exonuclease I (GE Healthcare) at 37°C for 40 min, followed by 20 min at 80°C. .. For multi-peak Vα and Vβ profiles, AV-AC and BV-BC PCR products were cloned using the TOPO TA cloning kit (Invitrogen) following the manufacturer's instructions.

Article Title: Population Genetic Structure of the Magnificent Frigatebird Fregata magnificens (Aves, Suliformes) Breeding Colonies in the Western Atlantic Ocean
Article Snippet: The PCR conditions were the same for the two mitochondrial segments: 10 cycles of 94°C for 60 s, 60°C for 60 s (-1μC/cycle), and 72°C for 60 s, followed by 35 cycles of 94°C for 60 s, 55°C for 60 s, and 72°C for 60 s, and a final extension of 72°C for 10 min. .. The amplified products were assessed on a 1% agarose gel, purified enzymatically with shrimp alkaline phosphatase and exonuclease I (GE Healthcare), and Sanger sequenced by Macrogen Inc., South Korea. .. We selected eight microsatellite (Short Tandem Repeat—STR) loci (Fmin02, Fmin11, Fmin12, Fmin14, Fmin15, Fmin16, Fmin17, and Fmin18; [ ]) developed for the Great Frigatebird, F . minor , which were shown to exhibit consistent amplification and polymorphism in F . magnificens [ ] Every forward primer was 5’-tailed with an M13 sequence ( 5’-CACGACGTTGTAAAACGAC-3’ ) and used in combination with an M13 primer that had the same sequence but was dye labelled at its 5’ end [ ].

Article Title: A Family of H723R Mutation for SLC26A4 Associated with Enlarged Vestibular Aqueduct Syndrome
Article Snippet: Amplification was carried out in a Perkin-Elmer thermal cycler using the following conditions: 5-min denaturation at 95℃, 37 three-step cycles (95℃ for 30 sec, 55℃ for 1 min, 72℃ for 1 or 3 min), 72℃ for 10 min, and ending with a holding period at 4℃. .. The PCR products were directly sequenced after removal of unincorporated dNTPs and primers by incubation of 50-100 ng PCR product at 37℃ for 30 min with 0.1 µL exonuclease I (Amersham Life Science, Cleveland, USA) and 1 µL shrimp alkaline phosphatase (American Life Science).

Polymerase Chain Reaction:

Article Title: The human squamous oesophagus has widespread capacity for clonal expansion from cells at diverse stages of differentiation
Article Snippet: Incubation conditions were as follows: 15 min at 50°C for RT, 2 min at 95°C for Taq inactivation, followed by a touch-down PCR amplification with 20 cycles of 15 s at 95°C for denaturation and 4 min for annealing and extension (decreasing temperature from 70°C down to 60°C over 5 cycles, followed by 15 cycles at 60°C) . .. Amplification was followed by DNA digestion with Exonuclease 1 (Exostar-1 step, GE Healthcare) as per the manufacturer's recommendation. qPCR was performed with LightCycler 480 SYBR Green I master (Roche Diagnostics Ltd, Rotkreuz, Switzerland) and 0.2 µM forward and reverse primer in a final volume of 10 µL using LightCycler 480 technology (Roche). qPCR consisted of 55 cycles of denaturation at 95°C (10 s), annealing at 60°C (15 s) and extension at 72°C (15 s).

Article Title: DNA damage predicts prognosis and treatment response in colorectal liver metastases superior to immunogenic cell death and T cells
Article Snippet: Briefly, genomic DNA was extracted from tissue blocks and exon 2 and 3 of the KRAS gene and exon 15 of the BRAF gene were polymerase chain reaction (PCR) amplified with AmpliTaq Gold® DNA polymerase (Applied Biosystems, Fisher Scientific, Vienna, Austria) and corresponding oligonucleotide primers ( Table ). .. Excess of primers and deoxynucleotides (dNTPs) was removed by incubation of 5 µL PCR product with 2.5 U exonuclease I (GE Healthcare Life Sciences, Vienna, Austria) and 2.5 U shrimp alkaline phosphatase (SAP; GE Healthcare Life Sciences) at 37°C for 1 h. Enzymes were further heat inactivated at 70°C for 15 min, following sequence analysis with 1-2 µL of the purified PCR product and 4 pmol primers using the BigDye™ Terminator Cycle Sequencing Kit (Applied Biosystems) according to the manufacturer's instructions. .. Excess of BigDye™ Terminator nucleotides was removed by centrifugation with Centri-Sep™ Spin Columns (Invitrogen, Fisher Scientific, Vienna, Austria).

Article Title: Ancient female philopatry, asymmetric male gene flow, and synchronous population expansion support the influence of climatic oscillations on the evolution of South American sea lion (Otaria flavescens)
Article Snippet: Amplifications for CR and cytb were carried out in 20 μl with the following conditions: 1.5 mM MgCl2, 200 μM of each dNTP, 0.1 μM of each primer, 1 U of Platinum Taq DNA polymerase (Invitrogen), 1X PCR buffer (Invitrogen), 0.2%–0.4% Triton and 2 μl of DNA (approximately 50 ng). .. Amplification products were purified with shrimp alkaline phosphatase and exonuclease I (Amersham Biosciences).

Article Title: Colorectal Tumors from APC*I1307K Carriers Principally Harbor Somatic APC Mutations outside the A8 Tract
Article Snippet: Samples in which a variant was identified by FSSCP were re-amplified using unlabeled PCR primers. .. PCR products were prepared for sequencing by treatment of 5 µl of PCR product with 2 U of shrimp alkaline phosphatase and 10 U of exonuclease I (Amersham Pharmacia, Chalfont St Giles, UK) at 37°C for 30 minutes and then 80°C for 15 minutes. .. Sequencing reactions of 10 µl were carried out using 1–3 µl of prepared PCR product, 1.6 picomoles of primer, BigDye ready reaction mix (Applied Biosystems) version 2 diluted 1∶2 with Half Big Dye reagent (Genetix, http://www.genetix.com/ ).

Article Title: Complete mitochondrial DNA sequence of the endangered fish (Bahabataipingensis): Mitogenome characterization and phylogenetic implications
Article Snippet: Negative controls were included in all PCR amplifications to confirm the absence of contaminants. .. PCR products were cleaned by adding 0.45 µl of Shrimp Alkaline Phosphatase (Biotech Pharmacon), 0.9 µl of Exonuclease I (GE Healthcare) and 1.65 µl of sterile distilled H2 O to 9 µL of PCR product and incubating at 37 °C for 30 min and 80 °C for 20 min. .. The purified product was then sequenced on ABI Prism 3730 (Applied Biosystems) from both strands with the same primers as those used for PCRs.

Article Title: Consuming viscous prey: a novel protein-secreting delivery system in neotropical snail-eating snakes
Article Snippet: Primers and PCR protocols for partial amplification of genes 12S, 16S, cytb, bdnf, and c-mos were those described in Grazziotin et al. [ ]. .. PCRs were purified with shrimp alkaline phosphatase and exonuclease I (GE Healthcare, Piscataway, NJ, USA) and sequences were processed using the DYEnamic ET Dye Terminator Cycle Sequencing Kit in a MegaBACE 1000 automated sequencer (GE Healthcare) following the manufacturer’s protocols.

Article Title: Specific variants in WDR35 cause a distinctive form of Ellis-van Creveld syndrome by disrupting the recruitment of the EvC complex and SMO into the cilium
Article Snippet: WDR 35 coding exons and at least 100 bp of the adjacent introns were amplified by standard PCR from peripheral blood genomic DNA. .. PCR products were enzymatically treated with alkaline phosphatase and exonuclease I (ExoSapit; GE Healthcare) prior to be sequenced using a dye terminator cycle sequencing kit and run on an ABI 3730 sequencer (Applied Biosystems). .. Sequencing of IFT 122 was conducted using an in-house next generation sequencing (NGS) panel for detection of pathogenic variants in skeletal dysplasias comprising 315 genes.

Article Title: Identification of a Mutation Causing Deficient BMP1/mTLD Proteolytic Activity in Autosomal Recessive Osteogenesis Imperfecta
Article Snippet: BMP1 coding exons and at least 100 bp of the flanking introns were amplified by standard PCR (primers available on request). .. Prior to sequencing, the PCR products were treated with shrimp alkaline phosphatase and exonuclease I (ExoSapit; GE Healthcare, Piscataway, NJ) according to the manufacturers instructions. .. Sequencing reactions were carried out using a dye terminator cycle sequencing kit and run on an ABI 3730 sequencer (Applied Biosystems, Foster City, CA).

Article Title: FGFR3 protein expression and its relationship to mutation status and prognostic variables in bladder cancer
Article Snippet: Primers used for PCR amplification have been described previously [ ]. .. For sequencing, unincorporated primers and deoxynucleotides were removed using shrimp alkaline phosphatase and exonuclease I (Amersham Biosciences, Chalfont, UK).

Article Title: Evolution of Smooth Tubercle Bacilli PE and PE_PGRS Genes: Evidence for a Prominent Role of Recombination and Imprint of Positive Selection
Article Snippet: Paragraph title: PCR amplification and DNA sequencing ... Amplicons were subjected to sequencing after treatment with Exonuclease I (Amersham Biosciences) and Shrimp Alkaline Phosphatase (Amersham Biosciences).

Article Title: Genetic Analyses of Heme Oxygenase 1 (HMOX1) in Different Forms of Pancreatitis
Article Snippet: Oligonucleotide sequences and annealing temperatures of the primers are listed in . .. We digested PCR products with shrimp alkaline phosphatase (USB) and exonuclease I (GE Healthcare) and performed cycle sequencing using BigDye terminator mix (Applied Biosystems). .. We purified reaction products with ethanol precipitation and loaded them onto an ABI 3100 fluorescence sequencer (Applied Biosystems).

Article Title: Association of Mitochondrial DNA Polymerase ? Gene POLG1 Polymorphisms with Parkinsonism in Chinese Populations
Article Snippet: Paragraph title: PCR and SNP screening by sequencing ... Before sequencing, amplification products were incubated with shrimp alkaline phosphatase (0.5 units; Amersham) and exonuclease I (1 units; Amersham) at 37°C for 30 min, followed by heat inactivation at 80°C for 15 min.

Article Title: Endogenous TCR Recombination in TCR Tg Single RAG-Deficient Mice Uncovered by Robust In Vivo T Cell Activation and Selection
Article Snippet: For single peak Vα and Vβ profiles, direct sequencing was performed with AV and AC, and BV and BC primers following the recommendations of the BigDye Terminator v1.1 Cycle Sequencing Ready Reaction kit (Applied Biosystems). .. PCR products were then incubated with 0.5 U of shrimp alkaline phosphatase and 5 U of exonuclease I (GE Healthcare) at 37°C for 40 min, followed by 20 min at 80°C. .. DNA alignments were performed using the GCG package (Genetics Computer Group, Accelrys, Cambridge, U.K.).

Article Title: Population Genetic Structure of the Magnificent Frigatebird Fregata magnificens (Aves, Suliformes) Breeding Colonies in the Western Atlantic Ocean
Article Snippet: The amplified products were assessed on a 1% agarose gel, purified enzymatically with shrimp alkaline phosphatase and exonuclease I (GE Healthcare), and Sanger sequenced by Macrogen Inc., South Korea. .. The amplified products were assessed on a 1% agarose gel, purified enzymatically with shrimp alkaline phosphatase and exonuclease I (GE Healthcare), and Sanger sequenced by Macrogen Inc., South Korea.

Article Title: A Family of H723R Mutation for SLC26A4 Associated with Enlarged Vestibular Aqueduct Syndrome
Article Snippet: Amplification was carried out in a Perkin-Elmer thermal cycler using the following conditions: 5-min denaturation at 95℃, 37 three-step cycles (95℃ for 30 sec, 55℃ for 1 min, 72℃ for 1 or 3 min), 72℃ for 10 min, and ending with a holding period at 4℃. .. The PCR products were directly sequenced after removal of unincorporated dNTPs and primers by incubation of 50-100 ng PCR product at 37℃ for 30 min with 0.1 µL exonuclease I (Amersham Life Science, Cleveland, USA) and 1 µL shrimp alkaline phosphatase (American Life Science). .. The enzymes were heat-inactivated at 80℃ for 15 min. An aliquot of 6 pmol of either the forward or the reverse primer was used in standard cycle sequencing reactions with ABI Big Dye terminators, and run on an ABI 377 sequencer.

Article Title: Ghrelin Gene Variants Influence on Metabolic Syndrome Components in Aged Spanish Population
Article Snippet: Products were treated with Exonuclease I (Amersham Biosciences) and shrimp alkaline phosphatase (Amersham Biosciences) to remove excess primers and deoxynucleotide triphosphates. .. Products were treated with Exonuclease I (Amersham Biosciences) and shrimp alkaline phosphatase (Amersham Biosciences) to remove excess primers and deoxynucleotide triphosphates.

Article Title: Mutation Spectrum of Meckel Syndrome Genes: One Group of Syndromes or Several Distinct Groups?
Article Snippet: Primer sequences are available upon request. .. The PCR reactions were done with AmpliTaq Gold and PCR products were treated with shrimp alkaline phosphatase (SAP) and Exonuclease I (Amersham) before sequencing. .. Sequencing was carried out using ABI BigDye chemistry with ABI3730 automated sequencer (Applied Biosystems, Darmstadt, Germany) and sequences were analyzed using Sequencher software (Gene Codes).

TA Cloning:

Article Title: Endogenous TCR Recombination in TCR Tg Single RAG-Deficient Mice Uncovered by Robust In Vivo T Cell Activation and Selection
Article Snippet: PCR products were then incubated with 0.5 U of shrimp alkaline phosphatase and 5 U of exonuclease I (GE Healthcare) at 37°C for 40 min, followed by 20 min at 80°C. .. PCR products were then incubated with 0.5 U of shrimp alkaline phosphatase and 5 U of exonuclease I (GE Healthcare) at 37°C for 40 min, followed by 20 min at 80°C.

Construct:

Article Title: Behavioural and Genetic Characteristics of a New Species of Nasonia
Article Snippet: To clean the reactions before sequencing, amplified reactions (8uL) were incubated with 0.5 U shrimp alkaline phosphatase and 1.0 U of exonuclease I (Amersham, Piscataway, NJ) along with the supplied buffer. .. To clean the reactions before sequencing, amplified reactions (8uL) were incubated with 0.5 U shrimp alkaline phosphatase and 1.0 U of exonuclease I (Amersham, Piscataway, NJ) along with the supplied buffer.

SYBR Green Assay:

Article Title: The human squamous oesophagus has widespread capacity for clonal expansion from cells at diverse stages of differentiation
Article Snippet: Incubation conditions were as follows: 15 min at 50°C for RT, 2 min at 95°C for Taq inactivation, followed by a touch-down PCR amplification with 20 cycles of 15 s at 95°C for denaturation and 4 min for annealing and extension (decreasing temperature from 70°C down to 60°C over 5 cycles, followed by 15 cycles at 60°C) . .. Amplification was followed by DNA digestion with Exonuclease 1 (Exostar-1 step, GE Healthcare) as per the manufacturer's recommendation. qPCR was performed with LightCycler 480 SYBR Green I master (Roche Diagnostics Ltd, Rotkreuz, Switzerland) and 0.2 µM forward and reverse primer in a final volume of 10 µL using LightCycler 480 technology (Roche). qPCR consisted of 55 cycles of denaturation at 95°C (10 s), annealing at 60°C (15 s) and extension at 72°C (15 s). .. Primer sequences are provided in online supplementary table S2.

Incubation:

Article Title: The human squamous oesophagus has widespread capacity for clonal expansion from cells at diverse stages of differentiation
Article Snippet: Incubation conditions were as follows: 15 min at 50°C for RT, 2 min at 95°C for Taq inactivation, followed by a touch-down PCR amplification with 20 cycles of 15 s at 95°C for denaturation and 4 min for annealing and extension (decreasing temperature from 70°C down to 60°C over 5 cycles, followed by 15 cycles at 60°C) . .. Amplification was followed by DNA digestion with Exonuclease 1 (Exostar-1 step, GE Healthcare) as per the manufacturer's recommendation. qPCR was performed with LightCycler 480 SYBR Green I master (Roche Diagnostics Ltd, Rotkreuz, Switzerland) and 0.2 µM forward and reverse primer in a final volume of 10 µL using LightCycler 480 technology (Roche). qPCR consisted of 55 cycles of denaturation at 95°C (10 s), annealing at 60°C (15 s) and extension at 72°C (15 s).

Article Title: DNA damage predicts prognosis and treatment response in colorectal liver metastases superior to immunogenic cell death and T cells
Article Snippet: Briefly, genomic DNA was extracted from tissue blocks and exon 2 and 3 of the KRAS gene and exon 15 of the BRAF gene were polymerase chain reaction (PCR) amplified with AmpliTaq Gold® DNA polymerase (Applied Biosystems, Fisher Scientific, Vienna, Austria) and corresponding oligonucleotide primers ( Table ). .. Excess of primers and deoxynucleotides (dNTPs) was removed by incubation of 5 µL PCR product with 2.5 U exonuclease I (GE Healthcare Life Sciences, Vienna, Austria) and 2.5 U shrimp alkaline phosphatase (SAP; GE Healthcare Life Sciences) at 37°C for 1 h. Enzymes were further heat inactivated at 70°C for 15 min, following sequence analysis with 1-2 µL of the purified PCR product and 4 pmol primers using the BigDye™ Terminator Cycle Sequencing Kit (Applied Biosystems) according to the manufacturer's instructions. .. Excess of BigDye™ Terminator nucleotides was removed by centrifugation with Centri-Sep™ Spin Columns (Invitrogen, Fisher Scientific, Vienna, Austria).

Article Title: Nanoliter high throughput quantitative PCR
Article Snippet: Cultures were incubated for 4 h at 37C, 5% CO2 . .. Samples were treated with Exonuclease I (10 U/μl) (Amersham Biosciences).

Article Title: Behavioural and Genetic Characteristics of a New Species of Nasonia
Article Snippet: NCBI accession numbers for each fragment from all the strains are summarized in . .. To clean the reactions before sequencing, amplified reactions (8uL) were incubated with 0.5 U shrimp alkaline phosphatase and 1.0 U of exonuclease I (Amersham, Piscataway, NJ) along with the supplied buffer. .. Sequencing was performed directly from the amplified products using a BigDye v3.0 terminator sequencing kit and an ABI 3700 or 3730 xl (Applied Biosystems, Foster City, CA) automated sequencer.

Article Title: Association of Mitochondrial DNA Polymerase ? Gene POLG1 Polymorphisms with Parkinsonism in Chinese Populations
Article Snippet: PCR conditions were set as follows: 2 min initial denaturation at 95°C, 30 cycles of 30 s denaturation at 94°C, 30 s annealing at 53–60°C, 30 s extension at 72°C and 10 min final extension at 72°C. .. Before sequencing, amplification products were incubated with shrimp alkaline phosphatase (0.5 units; Amersham) and exonuclease I (1 units; Amersham) at 37°C for 30 min, followed by heat inactivation at 80°C for 15 min. .. The DNA samples containing extension products were analyzed using the sequencing kit ABI PRISM dye terminator cycle and the automatic sequencing system ABI 3100 (Applied Biosystems).

Article Title: Endogenous TCR Recombination in TCR Tg Single RAG-Deficient Mice Uncovered by Robust In Vivo T Cell Activation and Selection
Article Snippet: For single peak Vα and Vβ profiles, direct sequencing was performed with AV and AC, and BV and BC primers following the recommendations of the BigDye Terminator v1.1 Cycle Sequencing Ready Reaction kit (Applied Biosystems). .. PCR products were then incubated with 0.5 U of shrimp alkaline phosphatase and 5 U of exonuclease I (GE Healthcare) at 37°C for 40 min, followed by 20 min at 80°C. .. DNA alignments were performed using the GCG package (Genetics Computer Group, Accelrys, Cambridge, U.K.).

Article Title: A Family of H723R Mutation for SLC26A4 Associated with Enlarged Vestibular Aqueduct Syndrome
Article Snippet: Amplification was carried out in a Perkin-Elmer thermal cycler using the following conditions: 5-min denaturation at 95℃, 37 three-step cycles (95℃ for 30 sec, 55℃ for 1 min, 72℃ for 1 or 3 min), 72℃ for 10 min, and ending with a holding period at 4℃. .. The PCR products were directly sequenced after removal of unincorporated dNTPs and primers by incubation of 50-100 ng PCR product at 37℃ for 30 min with 0.1 µL exonuclease I (Amersham Life Science, Cleveland, USA) and 1 µL shrimp alkaline phosphatase (American Life Science). .. The enzymes were heat-inactivated at 80℃ for 15 min. An aliquot of 6 pmol of either the forward or the reverse primer was used in standard cycle sequencing reactions with ABI Big Dye terminators, and run on an ABI 377 sequencer.

Expressing:

Article Title: The human squamous oesophagus has widespread capacity for clonal expansion from cells at diverse stages of differentiation
Article Snippet: Amplification was followed by DNA digestion with Exonuclease 1 (Exostar-1 step, GE Healthcare) as per the manufacturer's recommendation. qPCR was performed with LightCycler 480 SYBR Green I master (Roche Diagnostics Ltd, Rotkreuz, Switzerland) and 0.2 µM forward and reverse primer in a final volume of 10 µL using LightCycler 480 technology (Roche). qPCR consisted of 55 cycles of denaturation at 95°C (10 s), annealing at 60°C (15 s) and extension at 72°C (15 s). .. Results were analysed with LightCycler 480 software V.1.5 (Roche).

Countercurrent Chromatography:

Article Title: Ancient female philopatry, asymmetric male gene flow, and synchronous population expansion support the influence of climatic oscillations on the evolution of South American sea lion (Otaria flavescens)
Article Snippet: The mitochondrial DNA control region (CR) and cytochrome b (cytb) gene were partially amplified by PCR using the following primers: L15926 5´- TCA AAG CTT ACA CCA GTC TTG TAA ACC—3´ [ ]; H16498 5´- CCT GAA GTA GGA ACC AGA TG—3´ [ ] for the control region and GLUDG-L 5′-TGA CTT GAA RAA CCA YCG TTG-3′ and CB2-H 5′- CCC TCA GAA TGA TAT TTG TCC TCA-3′ [ ] for the cytb. .. Amplification products were purified with shrimp alkaline phosphatase and exonuclease I (Amersham Biosciences).

Flow Cytometry:

Article Title: The human squamous oesophagus has widespread capacity for clonal expansion from cells at diverse stages of differentiation
Article Snippet: Briefly, for each reaction 100 flow-sorted cells were incubated with 0.2 µL SuperScript III RT Platinum Taq Mix and 2.5 µL of primer mix in a final volume of 9 µL. .. Amplification was followed by DNA digestion with Exonuclease 1 (Exostar-1 step, GE Healthcare) as per the manufacturer's recommendation. qPCR was performed with LightCycler 480 SYBR Green I master (Roche Diagnostics Ltd, Rotkreuz, Switzerland) and 0.2 µM forward and reverse primer in a final volume of 10 µL using LightCycler 480 technology (Roche). qPCR consisted of 55 cycles of denaturation at 95°C (10 s), annealing at 60°C (15 s) and extension at 72°C (15 s).

Gas Chromatography:

Article Title: Evolution of Smooth Tubercle Bacilli PE and PE_PGRS Genes: Evidence for a Prominent Role of Recombination and Imprint of Positive Selection
Article Snippet: Although the genome sequence of some reference strains is publicly available, we PCR amplified and sequenced the PE/PE_PGRS selected members from the DNA of all these strains, since the accuracy of genome sequences in these highly GC-rich and repetitive regions is questioned. .. Amplicons were subjected to sequencing after treatment with Exonuclease I (Amersham Biosciences) and Shrimp Alkaline Phosphatase (Amersham Biosciences).

Genomic Sequencing:

Article Title: Identification of a Mutation Causing Deficient BMP1/mTLD Proteolytic Activity in Autosomal Recessive Osteogenesis Imperfecta
Article Snippet: Paragraph title: Genomic Sequencing ... Prior to sequencing, the PCR products were treated with shrimp alkaline phosphatase and exonuclease I (ExoSapit; GE Healthcare, Piscataway, NJ) according to the manufacturers instructions.

Generated:

Article Title: Behavioural and Genetic Characteristics of a New Species of Nasonia
Article Snippet: To clean the reactions before sequencing, amplified reactions (8uL) were incubated with 0.5 U shrimp alkaline phosphatase and 1.0 U of exonuclease I (Amersham, Piscataway, NJ) along with the supplied buffer. .. Sequencing was performed directly from the amplified products using a BigDye v3.0 terminator sequencing kit and an ABI 3700 or 3730 xl (Applied Biosystems, Foster City, CA) automated sequencer.

DNA Sequencing:

Article Title: Colorectal Tumors from APC*I1307K Carriers Principally Harbor Somatic APC Mutations outside the A8 Tract
Article Snippet: Paragraph title: Extended DNA Sequencing ... PCR products were prepared for sequencing by treatment of 5 µl of PCR product with 2 U of shrimp alkaline phosphatase and 10 U of exonuclease I (Amersham Pharmacia, Chalfont St Giles, UK) at 37°C for 30 minutes and then 80°C for 15 minutes.

Article Title: Evolution of Smooth Tubercle Bacilli PE and PE_PGRS Genes: Evidence for a Prominent Role of Recombination and Imprint of Positive Selection
Article Snippet: Paragraph title: PCR amplification and DNA sequencing ... Amplicons were subjected to sequencing after treatment with Exonuclease I (Amersham Biosciences) and Shrimp Alkaline Phosphatase (Amersham Biosciences).

Article Title: Genetic Analyses of Heme Oxygenase 1 (HMOX1) in Different Forms of Pancreatitis
Article Snippet: Paragraph title: Polymerase Chain Reaction and DNA sequencing ... We digested PCR products with shrimp alkaline phosphatase (USB) and exonuclease I (GE Healthcare) and performed cycle sequencing using BigDye terminator mix (Applied Biosystems).

Reverse Transcription Polymerase Chain Reaction:

Article Title: The human squamous oesophagus has widespread capacity for clonal expansion from cells at diverse stages of differentiation
Article Snippet: Reverse transcription (RT) and cDNA amplification were carried out in one step using the SuperScript III One-Step RT-PCR System (Invitrogen). .. Amplification was followed by DNA digestion with Exonuclease 1 (Exostar-1 step, GE Healthcare) as per the manufacturer's recommendation. qPCR was performed with LightCycler 480 SYBR Green I master (Roche Diagnostics Ltd, Rotkreuz, Switzerland) and 0.2 µM forward and reverse primer in a final volume of 10 µL using LightCycler 480 technology (Roche). qPCR consisted of 55 cycles of denaturation at 95°C (10 s), annealing at 60°C (15 s) and extension at 72°C (15 s).

Recombinant:

Article Title: Nanoliter high throughput quantitative PCR
Article Snippet: Recombinant Human TNF-α (10 ng/ml in EGM™-2), purchased from R & D Systems Inc. (Minneapolis, MN) was added to HUVEC cultures that were ∼50% confluent in T150 tissue culture flasks. .. Samples were treated with Exonuclease I (10 U/μl) (Amersham Biosciences).

DNA Extraction:

Article Title: FGFR3 protein expression and its relationship to mutation status and prognostic variables in bladder cancer
Article Snippet: Paragraph title: DNA extraction and FGFR3 mutation analysis ... For sequencing, unincorporated primers and deoxynucleotides were removed using shrimp alkaline phosphatase and exonuclease I (Amersham Biosciences, Chalfont, UK).

Mutagenesis:

Article Title: DNA damage predicts prognosis and treatment response in colorectal liver metastases superior to immunogenic cell death and T cells
Article Snippet: Paragraph title: Analysis of mutation status ... Excess of primers and deoxynucleotides (dNTPs) was removed by incubation of 5 µL PCR product with 2.5 U exonuclease I (GE Healthcare Life Sciences, Vienna, Austria) and 2.5 U shrimp alkaline phosphatase (SAP; GE Healthcare Life Sciences) at 37°C for 1 h. Enzymes were further heat inactivated at 70°C for 15 min, following sequence analysis with 1-2 µL of the purified PCR product and 4 pmol primers using the BigDye™ Terminator Cycle Sequencing Kit (Applied Biosystems) according to the manufacturer's instructions.

Article Title: Identification of a Mutation Causing Deficient BMP1/mTLD Proteolytic Activity in Autosomal Recessive Osteogenesis Imperfecta
Article Snippet: Prior to sequencing, the PCR products were treated with shrimp alkaline phosphatase and exonuclease I (ExoSapit; GE Healthcare, Piscataway, NJ) according to the manufacturers instructions. .. Chromatograms were aligned and compared with the mRNA reference nucleotide sequence of BMP1 ( and ) and with the corresponding genomic sequence obtained from the Ensembl genome browser using Sequencher (Gene Codes Corp, Ann Arbor, MI).

Article Title: FGFR3 protein expression and its relationship to mutation status and prognostic variables in bladder cancer
Article Snippet: Paragraph title: DNA extraction and FGFR3 mutation analysis ... For sequencing, unincorporated primers and deoxynucleotides were removed using shrimp alkaline phosphatase and exonuclease I (Amersham Biosciences, Chalfont, UK).

Article Title: Genetic Analyses of Heme Oxygenase 1 (HMOX1) in Different Forms of Pancreatitis
Article Snippet: We digested PCR products with shrimp alkaline phosphatase (USB) and exonuclease I (GE Healthcare) and performed cycle sequencing using BigDye terminator mix (Applied Biosystems). .. We purified reaction products with ethanol precipitation and loaded them onto an ABI 3100 fluorescence sequencer (Applied Biosystems).

Article Title: Mutation Spectrum of Meckel Syndrome Genes: One Group of Syndromes or Several Distinct Groups?
Article Snippet: Paragraph title: Mutation Analysis ... The PCR reactions were done with AmpliTaq Gold and PCR products were treated with shrimp alkaline phosphatase (SAP) and Exonuclease I (Amersham) before sequencing.

Isolation:

Article Title: Ghrelin Gene Variants Influence on Metabolic Syndrome Components in Aged Spanish Population
Article Snippet: DNA was isolated from peripheral blood cells using the Chemagic System (Chemagen; Baesweiler, Germany). .. Products were treated with Exonuclease I (Amersham Biosciences) and shrimp alkaline phosphatase (Amersham Biosciences) to remove excess primers and deoxynucleotide triphosphates.

Size-exclusion Chromatography:

Article Title: A Family of H723R Mutation for SLC26A4 Associated with Enlarged Vestibular Aqueduct Syndrome
Article Snippet: Amplification was carried out in a Perkin-Elmer thermal cycler using the following conditions: 5-min denaturation at 95℃, 37 three-step cycles (95℃ for 30 sec, 55℃ for 1 min, 72℃ for 1 or 3 min), 72℃ for 10 min, and ending with a holding period at 4℃. .. The PCR products were directly sequenced after removal of unincorporated dNTPs and primers by incubation of 50-100 ng PCR product at 37℃ for 30 min with 0.1 µL exonuclease I (Amersham Life Science, Cleveland, USA) and 1 µL shrimp alkaline phosphatase (American Life Science).

Purification:

Article Title: DNA damage predicts prognosis and treatment response in colorectal liver metastases superior to immunogenic cell death and T cells
Article Snippet: Briefly, genomic DNA was extracted from tissue blocks and exon 2 and 3 of the KRAS gene and exon 15 of the BRAF gene were polymerase chain reaction (PCR) amplified with AmpliTaq Gold® DNA polymerase (Applied Biosystems, Fisher Scientific, Vienna, Austria) and corresponding oligonucleotide primers ( Table ). .. Excess of primers and deoxynucleotides (dNTPs) was removed by incubation of 5 µL PCR product with 2.5 U exonuclease I (GE Healthcare Life Sciences, Vienna, Austria) and 2.5 U shrimp alkaline phosphatase (SAP; GE Healthcare Life Sciences) at 37°C for 1 h. Enzymes were further heat inactivated at 70°C for 15 min, following sequence analysis with 1-2 µL of the purified PCR product and 4 pmol primers using the BigDye™ Terminator Cycle Sequencing Kit (Applied Biosystems) according to the manufacturer's instructions. .. Excess of BigDye™ Terminator nucleotides was removed by centrifugation with Centri-Sep™ Spin Columns (Invitrogen, Fisher Scientific, Vienna, Austria).

Article Title: Ancient female philopatry, asymmetric male gene flow, and synchronous population expansion support the influence of climatic oscillations on the evolution of South American sea lion (Otaria flavescens)
Article Snippet: Amplifications for CR and cytb were carried out in 20 μl with the following conditions: 1.5 mM MgCl2, 200 μM of each dNTP, 0.1 μM of each primer, 1 U of Platinum Taq DNA polymerase (Invitrogen), 1X PCR buffer (Invitrogen), 0.2%–0.4% Triton and 2 μl of DNA (approximately 50 ng). .. Amplification products were purified with shrimp alkaline phosphatase and exonuclease I (Amersham Biosciences). .. The purified products were sequenced in both directions using the DYEnamic ET Dye Terminator Cycle Sequencing Kit (Amersham Biosciences) and run in a MegaBace 1000 automated sequencer (Amersham Biosciences).

Article Title: Nanoliter high throughput quantitative PCR
Article Snippet: RNA was purified using an RNeasy Mini Kit (Qiagen Inc.) and reverse transcription was done using a High Capacity cDNA Archive Kit (Applied Biosystems). .. Samples were treated with Exonuclease I (10 U/μl) (Amersham Biosciences).

Article Title: Consuming viscous prey: a novel protein-secreting delivery system in neotropical snail-eating snakes
Article Snippet: We used the primers and protocols described in Vidal and Hedges [ ], Noonan and Chippindale [ ], and Chiari et al. [ ] to amplify fragments for the nuclear genes jun, nt3, and rag1, respectively. .. PCRs were purified with shrimp alkaline phosphatase and exonuclease I (GE Healthcare, Piscataway, NJ, USA) and sequences were processed using the DYEnamic ET Dye Terminator Cycle Sequencing Kit in a MegaBACE 1000 automated sequencer (GE Healthcare) following the manufacturer’s protocols. .. Both strands were sequenced for all fragments and sequences were assembled using Geneious 5.5 [ ].

Article Title: Population Genetic Structure of the Magnificent Frigatebird Fregata magnificens (Aves, Suliformes) Breeding Colonies in the Western Atlantic Ocean
Article Snippet: The PCR conditions were the same for the two mitochondrial segments: 10 cycles of 94°C for 60 s, 60°C for 60 s (-1μC/cycle), and 72°C for 60 s, followed by 35 cycles of 94°C for 60 s, 55°C for 60 s, and 72°C for 60 s, and a final extension of 72°C for 10 min. .. The amplified products were assessed on a 1% agarose gel, purified enzymatically with shrimp alkaline phosphatase and exonuclease I (GE Healthcare), and Sanger sequenced by Macrogen Inc., South Korea. .. We selected eight microsatellite (Short Tandem Repeat—STR) loci (Fmin02, Fmin11, Fmin12, Fmin14, Fmin15, Fmin16, Fmin17, and Fmin18; [ ]) developed for the Great Frigatebird, F . minor , which were shown to exhibit consistent amplification and polymorphism in F . magnificens [ ] Every forward primer was 5’-tailed with an M13 sequence ( 5’-CACGACGTTGTAAAACGAC-3’ ) and used in combination with an M13 primer that had the same sequence but was dye labelled at its 5’ end [ ].

Article Title: Ghrelin Gene Variants Influence on Metabolic Syndrome Components in Aged Spanish Population
Article Snippet: Products were treated with Exonuclease I (Amersham Biosciences) and shrimp alkaline phosphatase (Amersham Biosciences) to remove excess primers and deoxynucleotide triphosphates. .. For the examination of the six SNPs, extension SNaPshot primers specific to the polymorphic sites were used for the SNaPshot minisequencing reaction using the ABI PRISM SNaPshot Multiplex Kit (Applied Biosystems).

Sequencing:

Article Title: DNA damage predicts prognosis and treatment response in colorectal liver metastases superior to immunogenic cell death and T cells
Article Snippet: Briefly, genomic DNA was extracted from tissue blocks and exon 2 and 3 of the KRAS gene and exon 15 of the BRAF gene were polymerase chain reaction (PCR) amplified with AmpliTaq Gold® DNA polymerase (Applied Biosystems, Fisher Scientific, Vienna, Austria) and corresponding oligonucleotide primers ( Table ). .. Excess of primers and deoxynucleotides (dNTPs) was removed by incubation of 5 µL PCR product with 2.5 U exonuclease I (GE Healthcare Life Sciences, Vienna, Austria) and 2.5 U shrimp alkaline phosphatase (SAP; GE Healthcare Life Sciences) at 37°C for 1 h. Enzymes were further heat inactivated at 70°C for 15 min, following sequence analysis with 1-2 µL of the purified PCR product and 4 pmol primers using the BigDye™ Terminator Cycle Sequencing Kit (Applied Biosystems) according to the manufacturer's instructions. .. Excess of BigDye™ Terminator nucleotides was removed by centrifugation with Centri-Sep™ Spin Columns (Invitrogen, Fisher Scientific, Vienna, Austria).

Article Title: Ancient female philopatry, asymmetric male gene flow, and synchronous population expansion support the influence of climatic oscillations on the evolution of South American sea lion (Otaria flavescens)
Article Snippet: Paragraph title: Mitochondrial DNA amplification, sequencing and analysis ... Amplification products were purified with shrimp alkaline phosphatase and exonuclease I (Amersham Biosciences).

Article Title: Colorectal Tumors from APC*I1307K Carriers Principally Harbor Somatic APC Mutations outside the A8 Tract
Article Snippet: Samples in which a variant was identified by FSSCP were re-amplified using unlabeled PCR primers. .. PCR products were prepared for sequencing by treatment of 5 µl of PCR product with 2 U of shrimp alkaline phosphatase and 10 U of exonuclease I (Amersham Pharmacia, Chalfont St Giles, UK) at 37°C for 30 minutes and then 80°C for 15 minutes. .. Sequencing reactions of 10 µl were carried out using 1–3 µl of prepared PCR product, 1.6 picomoles of primer, BigDye ready reaction mix (Applied Biosystems) version 2 diluted 1∶2 with Half Big Dye reagent (Genetix, http://www.genetix.com/ ).

Article Title: Consuming viscous prey: a novel protein-secreting delivery system in neotropical snail-eating snakes
Article Snippet: We used the primers and protocols described in Vidal and Hedges [ ], Noonan and Chippindale [ ], and Chiari et al. [ ] to amplify fragments for the nuclear genes jun, nt3, and rag1, respectively. .. PCRs were purified with shrimp alkaline phosphatase and exonuclease I (GE Healthcare, Piscataway, NJ, USA) and sequences were processed using the DYEnamic ET Dye Terminator Cycle Sequencing Kit in a MegaBACE 1000 automated sequencer (GE Healthcare) following the manufacturer’s protocols. .. Both strands were sequenced for all fragments and sequences were assembled using Geneious 5.5 [ ].

Article Title: Behavioural and Genetic Characteristics of a New Species of Nasonia
Article Snippet: NCBI accession numbers for each fragment from all the strains are summarized in . .. To clean the reactions before sequencing, amplified reactions (8uL) were incubated with 0.5 U shrimp alkaline phosphatase and 1.0 U of exonuclease I (Amersham, Piscataway, NJ) along with the supplied buffer. .. Sequencing was performed directly from the amplified products using a BigDye v3.0 terminator sequencing kit and an ABI 3700 or 3730 xl (Applied Biosystems, Foster City, CA) automated sequencer.

Article Title: Specific variants in WDR35 cause a distinctive form of Ellis-van Creveld syndrome by disrupting the recruitment of the EvC complex and SMO into the cilium
Article Snippet: WDR 35 coding exons and at least 100 bp of the adjacent introns were amplified by standard PCR from peripheral blood genomic DNA. .. PCR products were enzymatically treated with alkaline phosphatase and exonuclease I (ExoSapit; GE Healthcare) prior to be sequenced using a dye terminator cycle sequencing kit and run on an ABI 3730 sequencer (Applied Biosystems). .. Sequencing of IFT 122 was conducted using an in-house next generation sequencing (NGS) panel for detection of pathogenic variants in skeletal dysplasias comprising 315 genes.

Article Title: Identification of a Mutation Causing Deficient BMP1/mTLD Proteolytic Activity in Autosomal Recessive Osteogenesis Imperfecta
Article Snippet: BMP1 coding exons and at least 100 bp of the flanking introns were amplified by standard PCR (primers available on request). .. Prior to sequencing, the PCR products were treated with shrimp alkaline phosphatase and exonuclease I (ExoSapit; GE Healthcare, Piscataway, NJ) according to the manufacturers instructions. .. Sequencing reactions were carried out using a dye terminator cycle sequencing kit and run on an ABI 3730 sequencer (Applied Biosystems, Foster City, CA).

Article Title: FGFR3 protein expression and its relationship to mutation status and prognostic variables in bladder cancer
Article Snippet: 2 μl samples of extracted DNA were amplified in 50 μl reactions using AmpliTaq Gold (Applied Biosystems, Foster City, CA, USA). .. For sequencing, unincorporated primers and deoxynucleotides were removed using shrimp alkaline phosphatase and exonuclease I (Amersham Biosciences, Chalfont, UK). .. Sequencing reactions were carried out using both the forward and reverse PCR primers with the BigDye Terminator V1.1 Cycle Sequencing Kit (Applied Biosystems).

Article Title: Evolution of Smooth Tubercle Bacilli PE and PE_PGRS Genes: Evidence for a Prominent Role of Recombination and Imprint of Positive Selection
Article Snippet: The amplification ended with a final elongation step of 7 min at 72°C. .. Amplicons were subjected to sequencing after treatment with Exonuclease I (Amersham Biosciences) and Shrimp Alkaline Phosphatase (Amersham Biosciences). .. The reaction consisted of 1.5 µl of BigDye terminator cycle sequencing reagents, 4 µl of BigDye terminator cycle sequencing buffer, 1 µl of 20 µM concentrations of primers, as well as sufficient UltraPURE Distilled DNase, RNase-Free Water (Gibco/Invitrogen) to make a 20-µl reaction.

Article Title: Genetic Analyses of Heme Oxygenase 1 (HMOX1) in Different Forms of Pancreatitis
Article Snippet: Oligonucleotide sequences and annealing temperatures of the primers are listed in . .. We digested PCR products with shrimp alkaline phosphatase (USB) and exonuclease I (GE Healthcare) and performed cycle sequencing using BigDye terminator mix (Applied Biosystems). .. We purified reaction products with ethanol precipitation and loaded them onto an ABI 3100 fluorescence sequencer (Applied Biosystems).

Article Title: Association of Mitochondrial DNA Polymerase ? Gene POLG1 Polymorphisms with Parkinsonism in Chinese Populations
Article Snippet: PCR conditions were set as follows: 2 min initial denaturation at 95°C, 30 cycles of 30 s denaturation at 94°C, 30 s annealing at 53–60°C, 30 s extension at 72°C and 10 min final extension at 72°C. .. Before sequencing, amplification products were incubated with shrimp alkaline phosphatase (0.5 units; Amersham) and exonuclease I (1 units; Amersham) at 37°C for 30 min, followed by heat inactivation at 80°C for 15 min. .. The DNA samples containing extension products were analyzed using the sequencing kit ABI PRISM dye terminator cycle and the automatic sequencing system ABI 3100 (Applied Biosystems).

Article Title: Endogenous TCR Recombination in TCR Tg Single RAG-Deficient Mice Uncovered by Robust In Vivo T Cell Activation and Selection
Article Snippet: Paragraph title: Sequencing analyses ... PCR products were then incubated with 0.5 U of shrimp alkaline phosphatase and 5 U of exonuclease I (GE Healthcare) at 37°C for 40 min, followed by 20 min at 80°C.

Article Title: A Family of H723R Mutation for SLC26A4 Associated with Enlarged Vestibular Aqueduct Syndrome
Article Snippet: Mutations were detected by a combination of intronic polymerase chain reaction (PCR) amplification, using primers flanking each exon, and sequencing of the amplification products. .. The PCR products were directly sequenced after removal of unincorporated dNTPs and primers by incubation of 50-100 ng PCR product at 37℃ for 30 min with 0.1 µL exonuclease I (Amersham Life Science, Cleveland, USA) and 1 µL shrimp alkaline phosphatase (American Life Science).

Article Title: Mutation Spectrum of Meckel Syndrome Genes: One Group of Syndromes or Several Distinct Groups?
Article Snippet: Primer sequences are available upon request. .. The PCR reactions were done with AmpliTaq Gold and PCR products were treated with shrimp alkaline phosphatase (SAP) and Exonuclease I (Amersham) before sequencing. .. Sequencing was carried out using ABI BigDye chemistry with ABI3730 automated sequencer (Applied Biosystems, Darmstadt, Germany) and sequences were analyzed using Sequencher software (Gene Codes).

Blocking Assay:

Article Title: FGFR3 protein expression and its relationship to mutation status and prognostic variables in bladder cancer
Article Snippet: For sequencing, unincorporated primers and deoxynucleotides were removed using shrimp alkaline phosphatase and exonuclease I (Amersham Biosciences, Chalfont, UK). .. For sequencing, unincorporated primers and deoxynucleotides were removed using shrimp alkaline phosphatase and exonuclease I (Amersham Biosciences, Chalfont, UK).

Sample Prep:

Article Title: Nanoliter high throughput quantitative PCR
Article Snippet: Paragraph title: Human umbilical vein endothelial cells (HUVEC sample preparation ... Samples were treated with Exonuclease I (10 U/μl) (Amersham Biosciences).

Plasmid Preparation:

Article Title: Endogenous TCR Recombination in TCR Tg Single RAG-Deficient Mice Uncovered by Robust In Vivo T Cell Activation and Selection
Article Snippet: PCR products were then incubated with 0.5 U of shrimp alkaline phosphatase and 5 U of exonuclease I (GE Healthcare) at 37°C for 40 min, followed by 20 min at 80°C. .. For multi-peak Vα and Vβ profiles, AV-AC and BV-BC PCR products were cloned using the TOPO TA cloning kit (Invitrogen) following the manufacturer's instructions.

Software:

Article Title: The human squamous oesophagus has widespread capacity for clonal expansion from cells at diverse stages of differentiation
Article Snippet: Amplification was followed by DNA digestion with Exonuclease 1 (Exostar-1 step, GE Healthcare) as per the manufacturer's recommendation. qPCR was performed with LightCycler 480 SYBR Green I master (Roche Diagnostics Ltd, Rotkreuz, Switzerland) and 0.2 µM forward and reverse primer in a final volume of 10 µL using LightCycler 480 technology (Roche). qPCR consisted of 55 cycles of denaturation at 95°C (10 s), annealing at 60°C (15 s) and extension at 72°C (15 s). .. Amplification was followed by DNA digestion with Exonuclease 1 (Exostar-1 step, GE Healthcare) as per the manufacturer's recommendation. qPCR was performed with LightCycler 480 SYBR Green I master (Roche Diagnostics Ltd, Rotkreuz, Switzerland) and 0.2 µM forward and reverse primer in a final volume of 10 µL using LightCycler 480 technology (Roche). qPCR consisted of 55 cycles of denaturation at 95°C (10 s), annealing at 60°C (15 s) and extension at 72°C (15 s).

Article Title: Specific variants in WDR35 cause a distinctive form of Ellis-van Creveld syndrome by disrupting the recruitment of the EvC complex and SMO into the cilium
Article Snippet: PCR products were enzymatically treated with alkaline phosphatase and exonuclease I (ExoSapit; GE Healthcare) prior to be sequenced using a dye terminator cycle sequencing kit and run on an ABI 3730 sequencer (Applied Biosystems). .. PCR products were enzymatically treated with alkaline phosphatase and exonuclease I (ExoSapit; GE Healthcare) prior to be sequenced using a dye terminator cycle sequencing kit and run on an ABI 3730 sequencer (Applied Biosystems).

Article Title: FGFR3 protein expression and its relationship to mutation status and prognostic variables in bladder cancer
Article Snippet: For sequencing, unincorporated primers and deoxynucleotides were removed using shrimp alkaline phosphatase and exonuclease I (Amersham Biosciences, Chalfont, UK). .. Sequencing reactions were carried out using both the forward and reverse PCR primers with the BigDye Terminator V1.1 Cycle Sequencing Kit (Applied Biosystems).

Article Title: Mutation Spectrum of Meckel Syndrome Genes: One Group of Syndromes or Several Distinct Groups?
Article Snippet: Primers for amplifying the genomic DNA were designed using Primer3 software ( ) and provided by Sigma-Genosys. .. The PCR reactions were done with AmpliTaq Gold and PCR products were treated with shrimp alkaline phosphatase (SAP) and Exonuclease I (Amersham) before sequencing.

Real-time Polymerase Chain Reaction:

Article Title: The human squamous oesophagus has widespread capacity for clonal expansion from cells at diverse stages of differentiation
Article Snippet: Incubation conditions were as follows: 15 min at 50°C for RT, 2 min at 95°C for Taq inactivation, followed by a touch-down PCR amplification with 20 cycles of 15 s at 95°C for denaturation and 4 min for annealing and extension (decreasing temperature from 70°C down to 60°C over 5 cycles, followed by 15 cycles at 60°C) . .. Amplification was followed by DNA digestion with Exonuclease 1 (Exostar-1 step, GE Healthcare) as per the manufacturer's recommendation. qPCR was performed with LightCycler 480 SYBR Green I master (Roche Diagnostics Ltd, Rotkreuz, Switzerland) and 0.2 µM forward and reverse primer in a final volume of 10 µL using LightCycler 480 technology (Roche). qPCR consisted of 55 cycles of denaturation at 95°C (10 s), annealing at 60°C (15 s) and extension at 72°C (15 s). .. Primer sequences are provided in online supplementary table S2.

Article Title: Nanoliter high throughput quantitative PCR
Article Snippet: RNA samples were converted to randomly primed first strand cDNA using High Capacity cDNA Archive Kit (Applied BioSystems, Foster City, CA). .. To reduce non-specific product formation during qPCR, the cDNA sample was heated to 75°C for 10 min to inactivate the reverse transcriptase; snap chilled on ice for 5 min, then treated 1 h with 1.3 U/μl Exonuclease I (Amersham Biosciences, Piscataway, NJ). .. The Exonuclease I is heat inactivated at 85°C for 10 min and the resulting cDNA solution is stored at −20C.

Functional Assay:

Article Title: Population Genetic Structure of the Magnificent Frigatebird Fregata magnificens (Aves, Suliformes) Breeding Colonies in the Western Atlantic Ocean
Article Snippet: [ ] using species-specific primers, strongly suggesting that we are studying the functional cytb copy. .. The amplified products were assessed on a 1% agarose gel, purified enzymatically with shrimp alkaline phosphatase and exonuclease I (GE Healthcare), and Sanger sequenced by Macrogen Inc., South Korea.

Agarose Gel Electrophoresis:

Article Title: Population Genetic Structure of the Magnificent Frigatebird Fregata magnificens (Aves, Suliformes) Breeding Colonies in the Western Atlantic Ocean
Article Snippet: The PCR conditions were the same for the two mitochondrial segments: 10 cycles of 94°C for 60 s, 60°C for 60 s (-1μC/cycle), and 72°C for 60 s, followed by 35 cycles of 94°C for 60 s, 55°C for 60 s, and 72°C for 60 s, and a final extension of 72°C for 10 min. .. The amplified products were assessed on a 1% agarose gel, purified enzymatically with shrimp alkaline phosphatase and exonuclease I (GE Healthcare), and Sanger sequenced by Macrogen Inc., South Korea. .. We selected eight microsatellite (Short Tandem Repeat—STR) loci (Fmin02, Fmin11, Fmin12, Fmin14, Fmin15, Fmin16, Fmin17, and Fmin18; [ ]) developed for the Great Frigatebird, F . minor , which were shown to exhibit consistent amplification and polymorphism in F . magnificens [ ] Every forward primer was 5’-tailed with an M13 sequence ( 5’-CACGACGTTGTAAAACGAC-3’ ) and used in combination with an M13 primer that had the same sequence but was dye labelled at its 5’ end [ ].

Electrophoresis:

Article Title: Ghrelin Gene Variants Influence on Metabolic Syndrome Components in Aged Spanish Population
Article Snippet: Products were treated with Exonuclease I (Amersham Biosciences) and shrimp alkaline phosphatase (Amersham Biosciences) to remove excess primers and deoxynucleotide triphosphates. .. Products were treated with Exonuclease I (Amersham Biosciences) and shrimp alkaline phosphatase (Amersham Biosciences) to remove excess primers and deoxynucleotide triphosphates.

Ethanol Precipitation:

Article Title: Colorectal Tumors from APC*I1307K Carriers Principally Harbor Somatic APC Mutations outside the A8 Tract
Article Snippet: PCR products were prepared for sequencing by treatment of 5 µl of PCR product with 2 U of shrimp alkaline phosphatase and 10 U of exonuclease I (Amersham Pharmacia, Chalfont St Giles, UK) at 37°C for 30 minutes and then 80°C for 15 minutes. .. PCR products were prepared for sequencing by treatment of 5 µl of PCR product with 2 U of shrimp alkaline phosphatase and 10 U of exonuclease I (Amersham Pharmacia, Chalfont St Giles, UK) at 37°C for 30 minutes and then 80°C for 15 minutes.

Sampling:

Article Title: Consuming viscous prey: a novel protein-secreting delivery system in neotropical snail-eating snakes
Article Snippet: Sixty-one Dipsadinae terminals composed the ingroup, representing an increase of 25 species in respect to the taxon sampling used by Pyron et al. [ ] and 31 species to the one used by Grazziotin et al. [ ]. .. PCRs were purified with shrimp alkaline phosphatase and exonuclease I (GE Healthcare, Piscataway, NJ, USA) and sequences were processed using the DYEnamic ET Dye Terminator Cycle Sequencing Kit in a MegaBACE 1000 automated sequencer (GE Healthcare) following the manufacturer’s protocols.

Variant Assay:

Article Title: Colorectal Tumors from APC*I1307K Carriers Principally Harbor Somatic APC Mutations outside the A8 Tract
Article Snippet: Samples in which a variant was identified by FSSCP were re-amplified using unlabeled PCR primers. .. PCR products were prepared for sequencing by treatment of 5 µl of PCR product with 2 U of shrimp alkaline phosphatase and 10 U of exonuclease I (Amersham Pharmacia, Chalfont St Giles, UK) at 37°C for 30 minutes and then 80°C for 15 minutes.

Article Title: Specific variants in WDR35 cause a distinctive form of Ellis-van Creveld syndrome by disrupting the recruitment of the EvC complex and SMO into the cilium
Article Snippet: Paragraph title: Sequence variant screening and nomenclature ... PCR products were enzymatically treated with alkaline phosphatase and exonuclease I (ExoSapit; GE Healthcare) prior to be sequenced using a dye terminator cycle sequencing kit and run on an ABI 3730 sequencer (Applied Biosystems).

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    GE Healthcare exonuclease i shrimp alkaline phosphatase
    Exonuclease I Shrimp Alkaline Phosphatase, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 90/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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