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PerkinElmer geneamp pcr 9700 thermal cycler
Geneamp Pcr 9700 Thermal Cycler, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 89/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 89 stars, based on 16 article reviews
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geneamp pcr 9700 thermal cycler - by Bioz Stars, 2020-01
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RNA Extraction:

Article Title: Divergent ancestral lineages of newfound hantaviruses harbored by phylogenetically related crocidurine shrew species in Korea
Article Snippet: Paragraph title: RNA extraction, cDNA synthesis and RT-PCR amplification ... Initial denaturation at 94°C for 2 min was followed by two cycles each of denaturation at 94°C for 30 sec, two-degree step-down annealing from 46°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 30 cycles of denaturation at 94°C for 30 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA) ( ).

Article Title: Complex History of Codiversification and Host Switching of a Newfound Soricid-Borne Orthohantavirus in North America
Article Snippet: Paragraph title: 2.2. RNA Extraction, cDNA Synthesis, and RT-PCR Amplification ... Initial denaturation was followed by touchdown cycling (two-degree step-down annealing from 48 °C to 38 °C for 40 sec) and elongation at 72 °C for 1 min, then 32 cycles of denaturation at 94 °C for 40 sec, annealing at 42 °C for 40 sec, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA, USA).

Article Title: Highly Divergent Genetic Variants of Soricid-Borne Altai Virus (Hantaviridae) in Eurasia Suggest Ancient Host-Switching Events
Article Snippet: Paragraph title: 2.2. RNA Extraction and RT-PCR Analysis ... Initial denaturation at 94 °C for 5 min was followed by two cycles each of denaturation at 94 °C for 40 s, two-degree step-down annealing from 48 to 38 °C for 40 s, and elongation at 72 °C for 1 min, then 32 cycles of denaturation at 94 °C for 40 s, annealing at 42 °C for 40 s, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA, USA).

Article Title: Molecular Phylogeny of Hantaviruses Harbored by Insectivorous Bats in C?te d'Ivoire and Vietnam
Article Snippet: Genome Detection and Sequencing Total RNA extraction from tissues, using the PureLink Micro-to-Midi total RNA purification kit (Invitrogen, San Diego, CA, USA), and cDNA synthesis, using the SuperScript III First-Strand Synthesis Systems (Invitrogen) with random hexamers, were performed as described previously [ , , ]. .. Initial denaturation at 94 °C for 2 min was followed by two cycles each of denaturation at 94 °C for 30 s, two-degree step-down annealing from 46 °C to 38 °C for 40 s, and elongation at 72 °C for 1 min, then 30 cycles of denaturation at 94 °C for 30 s, annealing at 42 °C for 40 s, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA, USA) [ , , , , ].

Article Title: Shared Ancestry between a Newfound Mole-Borne Hantavirus and Hantaviruses Harbored by Cricetid Rodents ▿Shared Ancestry between a Newfound Mole-Borne Hantavirus and Hantaviruses Harbored by Cricetid Rodents ▿ †
Article Snippet: Paragraph title: RNA extraction and reverse transcription (RT)-PCR analysis. ... Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 s, 2°C step-down annealing from 48°C to 38°C for 40 s, and elongation at 72°C for 1 min and then 32 cycles of denaturation at 94°C for 40 s, annealing at 42°C for 40 s, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Sequencing:

Article Title: Complete genome sequence and molecular phylogeny of a newfound hantavirus harbored by the Doucet’s musk shrew (Crocidura douceti) in Guinea
Article Snippet: Innumerable trial-and-error testing of primers was necessary before ultimately obtaining the complete genome sequence of a newfound hantavirus ( ). .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Article Title: Isolation and partial characterization of a highly divergent lineage of hantavirus from the European mole (Talpa europaea)
Article Snippet: Paragraph title: Whole genome sequencing ... Initial denaturation at 94 °C for 2 min was followed by two cycles each of denaturation at 94 °C for 30 sec, two-degree step-down annealing from 46 °C to 38 °C for 40 sec, and elongation at 72 °C for 1 min, then 30 cycles of denaturation at 94 °C for 30 sec, annealing at 42 °C for 40 sec, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA) .

Article Title: Molecular Phylogeny of Hantaviruses Harbored by Insectivorous Bats in C?te d'Ivoire and Vietnam
Article Snippet: Paragraph title: 3.2. Genome Detection and Sequencing ... Initial denaturation at 94 °C for 2 min was followed by two cycles each of denaturation at 94 °C for 30 s, two-degree step-down annealing from 46 °C to 38 °C for 40 s, and elongation at 72 °C for 1 min, then 30 cycles of denaturation at 94 °C for 30 s, annealing at 42 °C for 40 s, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA, USA) [ , , , , ].

Article Title: Evolutionary Insights from a Genetically Divergent Hantavirus Harbored by the European Common Mole (Talpa europaea)
Article Snippet: RT-PCR and DNA Sequencing Initially, S- and L-segment sequences of rodent-borne hantaviruses, available in GenBank, and our growing sequence database of shrew-borne hantaviruses were aligned using ClustalW , and regions exhibiting high homology or sequence conservation were selected for designing oligonucleotide primers in nested RT-PCR. .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Agarose Gel Electrophoresis:

Article Title: Genetic basis for retention of a critical virulence plasmid of Borrelia burgdorferi
Article Snippet: The PCR conditions were 94°C for 2 min, followed by 30 cycles of 94°C for 45 s, 55°C for 45 s and 68°C for 3 min in a GeneAmp PCR 9700 thermal cycler (Perkin Elmer) or a DNA engine tetrad 2 thermal cycler (MJ Research). .. PCR products were separated by agarose gel electrophoresis and visualized by ethidium bromide staining.

Plasmid Preparation:

Article Title: Genetic basis for retention of a critical virulence plasmid of Borrelia burgdorferi
Article Snippet: PCR amplification with primers specific for the kanamycin resistance cassette (primers 48 and 49) was used to identify shuttle vector transformants. .. The PCR conditions were 94°C for 2 min, followed by 30 cycles of 94°C for 45 s, 55°C for 45 s and 68°C for 3 min in a GeneAmp PCR 9700 thermal cycler (Perkin Elmer) or a DNA engine tetrad 2 thermal cycler (MJ Research).

Synthesized:

Article Title: Divergent ancestral lineages of newfound hantaviruses harbored by phylogenetically related crocidurine shrew species in Korea
Article Snippet: Total RNA was extracted from 20–50 mg of each tissue, using the PureLink Micro-to-Midi total RNA purification kit (Invitrogen, San Diego, CA). cDNA, synthesized using the SuperScript III First-Strand Synthesis Systems (Invitrogen), were analyzed for hantavirus RNA by RT-PCR, using primers designed from highly conserved regions of hantavirus genomes ( , , ; , , ; , , ). .. Initial denaturation at 94°C for 2 min was followed by two cycles each of denaturation at 94°C for 30 sec, two-degree step-down annealing from 46°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 30 cycles of denaturation at 94°C for 30 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA) ( ).

Article Title: Genetic variants of Cao Bang hantavirus in the Chinese mole shrew (Anourosorex squamipes) and Taiwanese mole shrew (Anourosorex yamashinai)
Article Snippet: Total RNA was extracted from tissues using the PureLink Micro-to-Midi total RNA purification kit (Invitrogen, San Diego, CA), and cDNA was synthesized using the SuperScript III First-Strand Synthesis Systems (Invitrogen) with a highly conserved primer and/or random hexamers by two-step RT-PCR cycles. .. Initial denaturation at 94°C for 2 min was followed by two cycles each of denaturation at 94°C for 30 sec, two-degree step-down annealing from 46°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 30 cycles of denaturation at 94°C for 30 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Article Title: Dahonggou Creek virus, a divergent lineage of hantavirus harbored by the long-tailed mole (Scaptonyx fusicaudus)
Article Snippet: Briefly, total RNA was extracted from tissues using the PureLink Micro-to-Midi total RNA purification kit (Invitrogen, San Diego, CA), and cDNA was synthesized using the SuperScript III First-Strand Synthesis Systems (Invitrogen) with a highly conserved primer and/or random hexamers by two-step RT-PCR cycles. .. Initial denaturation at 94 °C for 2 min was followed by two cycles each of denaturation at 94 °C for 30 s, two-degree step-down annealing from 46 to 38 °C for 40 s, and elongation at 72 °C for 1 min, then 30 cycles of denaturation at 94 °C for 30 s, annealing at 42 °C for 40 s, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (PerkinElmer, Waltham, MA).

Article Title: Novel Hantavirus in the Flat-Skulled Shrew (Sorex roboratus)
Article Snippet: In an attempt to corroborate decades-old reports of hantaviral antigens in shrews from Russia (Gavrilovskaya et al. , Tkachenko et al. , Miasnikov et al. ), archival liver or lung tissues from 4 Siberian large-toothed shrews ( Sorex daphaenodon ), 5 Eurasian least shrews ( Sorex minutissimus ), 12 flat-skulled shrews ( Sorex roboratus ), and 18 tundra shrews ( Sorex tundrensis ), collected as part of the Beringian Coevolution Project in the Sakha Republic in northeastern Siberia ( ) during July and August 2006, were analyzed for hantavirus RNA by reverse transcription–polymerase chain reaction (PCR), using primers designed from highly conserved regions of hantavirus genomes (Arai et al. , Kang et al. , ). cDNA was synthesized using the SuperScript III First-Strand Synthesis System (Invitrogen, San Diego, CA) and an oligonucleotide primer (OSM55, 5′-TAGTAGTAGACTCC-3′) designed from the conserved 3′-end of the S, M, and L segments of hantaviruses. .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 s, two-degree step-down annealing from 48°C to 38°C for 40 s, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 s, annealing at 42°C for 40 s, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Isolation:

Article Title: Genetic basis for retention of a critical virulence plasmid of Borrelia burgdorferi
Article Snippet: The PCR conditions were 94°C for 2 min, followed by 30 cycles of 94°C for 45 s, 55°C for 45 s and 68°C for 3 min in a GeneAmp PCR 9700 thermal cycler (Perkin Elmer) or a DNA engine tetrad 2 thermal cycler (MJ Research). .. Total genomic DNA was then isolated from these cultures with a Wizard genomic DNA purification kit (Promega).

Genomic Sequencing:

Article Title: Divergent lineage of a novel hantavirus in the banana pipistrelle (Neoromicia nanus) in C?te d'Ivoire
Article Snippet: Using the PureLink Micro-to-Midi total RNA purification kit (Invitrogen, San Diego, CA), total RNA was extracted from 168 frozen and 45 ethanol-fixed liver and other visceral tissues of 213 insectivorous bats (representing 13 genera), collected during May 1981 to June 2011 in Asia, Africa and the Americas (Table ). cDNA was then prepared with the SuperScript III First-Strand Synthesis System (Invitrogen) using random hexamers, and PCR was performed as described previously, using an extensive panel of oligonucleotide primers, designed on conserved genomic sequences of rodent- and soricomorph-borne hantaviruses [ - , , ]. .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 s, two-degree step-down annealing from 48°C to 38°C for 40 s, and elongation at 72°C for 1 min or 1 min 20 s, then 32 cycles of denaturation at 94°C for 40 s, annealing at 42°C for 40 s, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Northern Blot:

Article Title: Novel Hantavirus in the Flat-Skulled Shrew (Sorex roboratus)
Article Snippet: Newly identified shrew-borne hantaviruses include Tanganya virus in the Therese's shrew ( Crocidura theresae ) (Klempa et al. ), Camp Ripley virus (RPLV) in the northern short-tailed shrew ( Blarina brevicauda ) (Arai et al. ), Cao Bang virus (CBNV) in the Chinese mole shrew ( Anourosorex squamipes ) (Song et al. ), Seewis virus (SWSV) in the Eurasian common shrew ( Sorex araneus ) (Song et al. ), Ash River virus (ARRV) in the masked shrew ( Sorex cinereus ) (Arai et al. ), Jemez Springs virus (JMSV) in the dusky shrew ( Sorex monticolus ) (Arai et al. ), and Imjin virus (MJNV) in the Ussuri white-toothed shrew ( Crocidura lasiura ) (Song et al. ). .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 s, two-degree step-down annealing from 48°C to 38°C for 40 s, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 s, annealing at 42°C for 40 s, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Size-exclusion Chromatography:

Article Title: Complete genome sequence and molecular phylogeny of a newfound hantavirus harbored by the Doucet’s musk shrew (Crocidura douceti) in Guinea
Article Snippet: .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany), and DNA sequencing was performed using an ABI Prism 377XL Genetic Analyzer (Applied Biosystems, Foster City, CA).

Article Title: Divergent ancestral lineages of newfound hantaviruses harbored by phylogenetically related crocidurine shrew species in Korea
Article Snippet: .. Initial denaturation at 94°C for 2 min was followed by two cycles each of denaturation at 94°C for 30 sec, two-degree step-down annealing from 46°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 30 cycles of denaturation at 94°C for 30 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA) ( ). .. PCR products were separated, using MobiSpin S-400 spin columns (MoBiTec, Goettingen, Germany), and amplicons were sequenced directly using an ABI Prism 3130 Genetic Analyzer (Applied Biosystems, Foster City, CA).

Article Title: Genetic variants of Cao Bang hantavirus in the Chinese mole shrew (Anourosorex squamipes) and Taiwanese mole shrew (Anourosorex yamashinai)
Article Snippet: .. Initial denaturation at 94°C for 2 min was followed by two cycles each of denaturation at 94°C for 30 sec, two-degree step-down annealing from 46°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 30 cycles of denaturation at 94°C for 30 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. PCR products were separated, using MobiSpin S-400 spin columns (MoBiTec, Goettingen, Germany), and amplicons were sequenced directly using an ABI Prism 3130 Genetic Analyzer (Applied Biosystems, Foster City, CA).

Article Title: Molecular evolution of Azagny virus, a newfound hantavirus harbored by the West African pygmy shrew (Crocidura obscurior) in C?te d'Ivoire
Article Snippet: .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany), and DNA sequencing was performed using an ABI Prism 377XL Genetic Analyzer (Applied Biosystems, Foster City, CA).

Article Title: Isolation and partial characterization of a highly divergent lineage of hantavirus from the European mole (Talpa europaea)
Article Snippet: .. Initial denaturation at 94 °C for 2 min was followed by two cycles each of denaturation at 94 °C for 30 sec, two-degree step-down annealing from 46 °C to 38 °C for 40 sec, and elongation at 72 °C for 1 min, then 30 cycles of denaturation at 94 °C for 30 sec, annealing at 42 °C for 40 sec, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA) . .. PCR products were separated, using MobiSpin S-400 spin columns (MoBiTec, Goettingen, Germany), and amplicons were sequenced directly using an ABI Prism 3130 Genetic Analyzer (Applied Biosystems, Foster City, CA) .

Article Title: Complex History of Codiversification and Host Switching of a Newfound Soricid-Borne Orthohantavirus in North America
Article Snippet: .. Initial denaturation was followed by touchdown cycling (two-degree step-down annealing from 48 °C to 38 °C for 40 sec) and elongation at 72 °C for 1 min, then 32 cycles of denaturation at 94 °C for 40 sec, annealing at 42 °C for 40 sec, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA, USA). .. Amplified products were separated by electrophoresis on 1.5% agarose gels and purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany).

Article Title: Evolutionary Insights from a Genetically Divergent Hantavirus Harbored by the European Common Mole (Talpa europaea)
Article Snippet: .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were separated by electrophoresis on 1.5% agarose gels and purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany).

Purification:

Article Title: Complete genome sequence and molecular phylogeny of a newfound hantavirus harbored by the Doucet’s musk shrew (Crocidura douceti) in Guinea
Article Snippet: Total RNA was extracted, using the PureLink Micro-to-Midi total RNA purification kit (Invitrogen, San Diego, CA, USA), from shrew tissues, and cDNA was prepared using the SuperScript III First-Strand Synthesis System (Invitrogen) and random hexamers and/or an oligonucleotide primer (OSM55: 5′-TAGTAGTAGACTCC-3′) designed from the conserved 5′-end of the S, M and L segments of hantaviruses. .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Article Title: Divergent ancestral lineages of newfound hantaviruses harbored by phylogenetically related crocidurine shrew species in Korea
Article Snippet: Total RNA was extracted from 20–50 mg of each tissue, using the PureLink Micro-to-Midi total RNA purification kit (Invitrogen, San Diego, CA). cDNA, synthesized using the SuperScript III First-Strand Synthesis Systems (Invitrogen), were analyzed for hantavirus RNA by RT-PCR, using primers designed from highly conserved regions of hantavirus genomes ( , , ; , , ; , , ). .. Initial denaturation at 94°C for 2 min was followed by two cycles each of denaturation at 94°C for 30 sec, two-degree step-down annealing from 46°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 30 cycles of denaturation at 94°C for 30 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA) ( ).

Article Title: Genetic variants of Cao Bang hantavirus in the Chinese mole shrew (Anourosorex squamipes) and Taiwanese mole shrew (Anourosorex yamashinai)
Article Snippet: Total RNA was extracted from tissues using the PureLink Micro-to-Midi total RNA purification kit (Invitrogen, San Diego, CA), and cDNA was synthesized using the SuperScript III First-Strand Synthesis Systems (Invitrogen) with a highly conserved primer and/or random hexamers by two-step RT-PCR cycles. .. Initial denaturation at 94°C for 2 min was followed by two cycles each of denaturation at 94°C for 30 sec, two-degree step-down annealing from 46°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 30 cycles of denaturation at 94°C for 30 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Article Title: Molecular evolution of Azagny virus, a newfound hantavirus harbored by the West African pygmy shrew (Crocidura obscurior) in C?te d'Ivoire
Article Snippet: Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany), and DNA sequencing was performed using an ABI Prism 377XL Genetic Analyzer (Applied Biosystems, Foster City, CA).

Article Title: Complex History of Codiversification and Host Switching of a Newfound Soricid-Borne Orthohantavirus in North America
Article Snippet: RNA Extraction, cDNA Synthesis, and RT-PCR Amplification Total RNA was extracted from 20–50 mg of each tissue using the PureLink Micro-to-Midi total RNA purification kit (Invitrogen, San Diego, CA, USA). cDNA was prepared using the SuperScript III First-Strand Synthesis System (Invitrogen) and oligonucleotide primer (5’-TAGTAGTAGACTCC-3’) designed from the conserved 3’-end of the S, M, and L segments of orthohantaviruses. .. Initial denaturation was followed by touchdown cycling (two-degree step-down annealing from 48 °C to 38 °C for 40 sec) and elongation at 72 °C for 1 min, then 32 cycles of denaturation at 94 °C for 40 sec, annealing at 42 °C for 40 sec, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA, USA).

Article Title: Divergent lineage of a novel hantavirus in the banana pipistrelle (Neoromicia nanus) in C?te d'Ivoire
Article Snippet: Using the PureLink Micro-to-Midi total RNA purification kit (Invitrogen, San Diego, CA), total RNA was extracted from 168 frozen and 45 ethanol-fixed liver and other visceral tissues of 213 insectivorous bats (representing 13 genera), collected during May 1981 to June 2011 in Asia, Africa and the Americas (Table ). cDNA was then prepared with the SuperScript III First-Strand Synthesis System (Invitrogen) using random hexamers, and PCR was performed as described previously, using an extensive panel of oligonucleotide primers, designed on conserved genomic sequences of rodent- and soricomorph-borne hantaviruses [ - , , ]. .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 s, two-degree step-down annealing from 48°C to 38°C for 40 s, and elongation at 72°C for 1 min or 1 min 20 s, then 32 cycles of denaturation at 94°C for 40 s, annealing at 42°C for 40 s, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Article Title: Highly Divergent Genetic Variants of Soricid-Borne Altai Virus (Hantaviridae) in Eurasia Suggest Ancient Host-Switching Events
Article Snippet: RNA Extraction and RT-PCR Analysis Total RNA was extracted from lung tissues, using the PureLink Micro-to-Midi total RNA purification kit (Invitrogen, San Diego, CA, USA), then reverse transcribed, using the SuperScript III First-Strand Synthesis System (Invitrogen) with random hexamers and universal oligonucleotide primer (OSM55, 5′-TAGTAGTAGACTCC-3′), designed from the conserved 3′ end of the S and L segments of hantaviruses [ , , , ]. .. Initial denaturation at 94 °C for 5 min was followed by two cycles each of denaturation at 94 °C for 40 s, two-degree step-down annealing from 48 to 38 °C for 40 s, and elongation at 72 °C for 1 min, then 32 cycles of denaturation at 94 °C for 40 s, annealing at 42 °C for 40 s, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA, USA).

Article Title: Dahonggou Creek virus, a divergent lineage of hantavirus harbored by the long-tailed mole (Scaptonyx fusicaudus)
Article Snippet: Briefly, total RNA was extracted from tissues using the PureLink Micro-to-Midi total RNA purification kit (Invitrogen, San Diego, CA), and cDNA was synthesized using the SuperScript III First-Strand Synthesis Systems (Invitrogen) with a highly conserved primer and/or random hexamers by two-step RT-PCR cycles. .. Initial denaturation at 94 °C for 2 min was followed by two cycles each of denaturation at 94 °C for 30 s, two-degree step-down annealing from 46 to 38 °C for 40 s, and elongation at 72 °C for 1 min, then 30 cycles of denaturation at 94 °C for 30 s, annealing at 42 °C for 40 s, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (PerkinElmer, Waltham, MA).

Article Title: Molecular Phylogeny of Hantaviruses Harbored by Insectivorous Bats in C?te d'Ivoire and Vietnam
Article Snippet: Genome Detection and Sequencing Total RNA extraction from tissues, using the PureLink Micro-to-Midi total RNA purification kit (Invitrogen, San Diego, CA, USA), and cDNA synthesis, using the SuperScript III First-Strand Synthesis Systems (Invitrogen) with random hexamers, were performed as described previously [ , , ]. .. Initial denaturation at 94 °C for 2 min was followed by two cycles each of denaturation at 94 °C for 30 s, two-degree step-down annealing from 46 °C to 38 °C for 40 s, and elongation at 72 °C for 1 min, then 30 cycles of denaturation at 94 °C for 30 s, annealing at 42 °C for 40 s, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA, USA) [ , , , , ].

Article Title: Novel Hantavirus in the Flat-Skulled Shrew (Sorex roboratus)
Article Snippet: Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 s, two-degree step-down annealing from 48°C to 38°C for 40 s, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 s, annealing at 42°C for 40 s, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were separated by electrophoresis on 1.5% agarose gels and purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany).

Article Title: Shared Ancestry between a Newfound Mole-Borne Hantavirus and Hantaviruses Harbored by Cricetid Rodents ▿Shared Ancestry between a Newfound Mole-Borne Hantavirus and Hantaviruses Harbored by Cricetid Rodents ▿ †
Article Snippet: Total RNA was extracted from tissues, using the PureLink Micro-to-Midi total RNA purification kit (Invitrogen, San Diego, CA), and then reverse transcribed, using the SuperScript III first-strand synthesis system (Invitrogen) and oligonucleotide primer (OSM55, 5′-TAGTAGTAGACTCC-3′), designed from the conserved 3′ ends of the S, M, and L segments of hantaviruses. .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 s, 2°C step-down annealing from 48°C to 38°C for 40 s, and elongation at 72°C for 1 min and then 32 cycles of denaturation at 94°C for 40 s, annealing at 42°C for 40 s, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Article Title: Evolutionary Insights from a Genetically Divergent Hantavirus Harbored by the European Common Mole (Talpa europaea)
Article Snippet: Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were separated by electrophoresis on 1.5% agarose gels and purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany).

Electrophoresis:

Article Title: Complex History of Codiversification and Host Switching of a Newfound Soricid-Borne Orthohantavirus in North America
Article Snippet: Initial denaturation was followed by touchdown cycling (two-degree step-down annealing from 48 °C to 38 °C for 40 sec) and elongation at 72 °C for 1 min, then 32 cycles of denaturation at 94 °C for 40 sec, annealing at 42 °C for 40 sec, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA, USA). .. Amplified products were separated by electrophoresis on 1.5% agarose gels and purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany).

Article Title: Highly Divergent Genetic Variants of Soricid-Borne Altai Virus (Hantaviridae) in Eurasia Suggest Ancient Host-Switching Events
Article Snippet: Initial denaturation at 94 °C for 5 min was followed by two cycles each of denaturation at 94 °C for 40 s, two-degree step-down annealing from 48 to 38 °C for 40 s, and elongation at 72 °C for 1 min, then 32 cycles of denaturation at 94 °C for 40 s, annealing at 42 °C for 40 s, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA, USA). .. Amplicons were separated by electrophoresis on 1.5% agarose gels and purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany).

Article Title: Novel Hantavirus in the Flat-Skulled Shrew (Sorex roboratus)
Article Snippet: Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 s, two-degree step-down annealing from 48°C to 38°C for 40 s, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 s, annealing at 42°C for 40 s, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were separated by electrophoresis on 1.5% agarose gels and purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany).

Article Title: Shared Ancestry between a Newfound Mole-Borne Hantavirus and Hantaviruses Harbored by Cricetid Rodents ▿Shared Ancestry between a Newfound Mole-Borne Hantavirus and Hantaviruses Harbored by Cricetid Rodents ▿ †
Article Snippet: Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 s, 2°C step-down annealing from 48°C to 38°C for 40 s, and elongation at 72°C for 1 min and then 32 cycles of denaturation at 94°C for 40 s, annealing at 42°C for 40 s, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were separated by electrophoresis on 1.5% agarose gels and purified using the QIAquick gel extraction kit (Qiagen, Hilden, Germany).

Article Title: Evolutionary Insights from a Genetically Divergent Hantavirus Harbored by the European Common Mole (Talpa europaea)
Article Snippet: Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were separated by electrophoresis on 1.5% agarose gels and purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany).

Modification:

Article Title: Complete genome sequence and molecular phylogeny of a newfound hantavirus harbored by the Doucet’s musk shrew (Crocidura douceti) in Guinea
Article Snippet: PCR was performed as described previously, with each 20-μL reaction containing 250 μM dNTP, 2 mM MgCl2 , 1 U of AmpliTaq polymerase (Roche, Basel, Switzerland) and 0.25 μM of oligonucleotide primers, with modified cycling conditions. .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Article Title: Molecular evolution of Azagny virus, a newfound hantavirus harbored by the West African pygmy shrew (Crocidura obscurior) in C?te d'Ivoire
Article Snippet: RT-PCR and DNA sequencing PCR was performed as described previously [ ], with each 20-μL reaction containing 250 μM dNTP, 2 mM MgCl2 , 1 U of AmpliTaq polymerase (Roche, Basel, Switzerland) and 0.25 μM of oligonucleotide primers, with trial-and-error testing of primers and modified cycling conditions. .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Generated:

Article Title: Genetic variants of Cao Bang hantavirus in the Chinese mole shrew (Anourosorex squamipes) and Taiwanese mole shrew (Anourosorex yamashinai)
Article Snippet: Initial denaturation at 94°C for 2 min was followed by two cycles each of denaturation at 94°C for 30 sec, two-degree step-down annealing from 46°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 30 cycles of denaturation at 94°C for 30 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Phylogenetic trees were generated by maximum likelihood and Bayesian methods, implemented in PAUP* (Phylogenetic Analysis Using Parsimony, 4.0b10) , RAxML Blackbox webserver ( ) and MrBayes 3.1 , under the best-fit GTR+I+Γ model of evolution selected by hierarchical likelihood-ratio test in MrModeltest v2.3 ( ) and jModelTest version 0.1 ( ).

Article Title: Dahonggou Creek virus, a divergent lineage of hantavirus harbored by the long-tailed mole (Scaptonyx fusicaudus)
Article Snippet: Initial denaturation at 94 °C for 2 min was followed by two cycles each of denaturation at 94 °C for 30 s, two-degree step-down annealing from 46 to 38 °C for 40 s, and elongation at 72 °C for 1 min, then 30 cycles of denaturation at 94 °C for 30 s, annealing at 42 °C for 40 s, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (PerkinElmer, Waltham, MA). .. Phylogenetic trees were generated using maximum likelihood and Bayesian methods, implemented in the RAxML Blackbox webserver [ ] and MrBayes 3.1 [ ], under the best-fit GTR+I+Γ model of evolution selected by hierarchical likelihood-ratio test in MrModeltest v2.3 [ ] and jModelTest version 0.1 [ ].

Article Title: Molecular Phylogenetic Analysis of Orientia tsutsugamushi Based on the groES and groEL Genes
Article Snippet: Initial denaturation was conducted at 94°C for 2 min, followed by 30 cycles of denaturation at 94°C for 40 s, annealing at 55°C for 30 s, and extension at 72°C for 2 min using a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. The phylogenetic analysis was conducted using Bayesian interference trees, which were generated with Bayesian Metropolis Hastings Markov Chain Monte Carlo (MCMC) tree-sampling methods implemented in MrBayes (Ronquist and Huelsenbeck ) using a GTR+I+Γ model of evolution, i.e ., the hierarchical likelihood-ratio test (hLRT) in MrModeltest2.3 ( ) (Posada and Crandall ) with partitioning based on codon positions.

Amplification:

Article Title: Divergent ancestral lineages of newfound hantaviruses harbored by phylogenetically related crocidurine shrew species in Korea
Article Snippet: Paragraph title: RNA extraction, cDNA synthesis and RT-PCR amplification ... Initial denaturation at 94°C for 2 min was followed by two cycles each of denaturation at 94°C for 30 sec, two-degree step-down annealing from 46°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 30 cycles of denaturation at 94°C for 30 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA) ( ).

Article Title: Isolation and partial characterization of a highly divergent lineage of hantavirus from the European mole (Talpa europaea)
Article Snippet: The 5′– and 3′–termini of each segment were amplified using the 3′–Full RACE Core Set (Takara Bio Inc., Otsu, Japan). .. Initial denaturation at 94 °C for 2 min was followed by two cycles each of denaturation at 94 °C for 30 sec, two-degree step-down annealing from 46 °C to 38 °C for 40 sec, and elongation at 72 °C for 1 min, then 30 cycles of denaturation at 94 °C for 30 sec, annealing at 42 °C for 40 sec, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA) .

Article Title: Genetic basis for retention of a critical virulence plasmid of Borrelia burgdorferi
Article Snippet: PCR amplification with primers specific for the kanamycin resistance cassette (primers 48 and 49) was used to identify shuttle vector transformants. .. The PCR conditions were 94°C for 2 min, followed by 30 cycles of 94°C for 45 s, 55°C for 45 s and 68°C for 3 min in a GeneAmp PCR 9700 thermal cycler (Perkin Elmer) or a DNA engine tetrad 2 thermal cycler (MJ Research).

Article Title: Complex History of Codiversification and Host Switching of a Newfound Soricid-Borne Orthohantavirus in North America
Article Snippet: Paragraph title: 2.2. RNA Extraction, cDNA Synthesis, and RT-PCR Amplification ... Initial denaturation was followed by touchdown cycling (two-degree step-down annealing from 48 °C to 38 °C for 40 sec) and elongation at 72 °C for 1 min, then 32 cycles of denaturation at 94 °C for 40 sec, annealing at 42 °C for 40 sec, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA, USA).

Article Title: Divergent lineage of a novel hantavirus in the banana pipistrelle (Neoromicia nanus) in C?te d'Ivoire
Article Snippet: Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 s, two-degree step-down annealing from 48°C to 38°C for 40 s, and elongation at 72°C for 1 min or 1 min 20 s, then 32 cycles of denaturation at 94°C for 40 s, annealing at 42°C for 40 s, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. The taxonomic identity of the hantavirus-infected vesper bats was confirmed by phylogenetic analysis of the cytochrome b gene of mtDNA (GenBank JQ287717 ), amplified by PCR as previously described [ , ].

Article Title: Highly Divergent Genetic Variants of Soricid-Borne Altai Virus (Hantaviridae) in Eurasia Suggest Ancient Host-Switching Events
Article Snippet: For the amplification of hantavirus genes, a two-step PCR was performed in 20-μL reaction mixtures, containing 250 μM dNTP, 2 mM MgCl2 , 1 U of AmpliTaq polymerase (Roche, Basel, Switzerland) and 0.25 μM of each oligonucleotide primer [ , , , ]. .. Initial denaturation at 94 °C for 5 min was followed by two cycles each of denaturation at 94 °C for 40 s, two-degree step-down annealing from 48 to 38 °C for 40 s, and elongation at 72 °C for 1 min, then 32 cycles of denaturation at 94 °C for 40 s, annealing at 42 °C for 40 s, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA, USA).

Article Title: Shared Ancestry between a Newfound Mole-Borne Hantavirus and Hantaviruses Harbored by Cricetid Rodents ▿Shared Ancestry between a Newfound Mole-Borne Hantavirus and Hantaviruses Harbored by Cricetid Rodents ▿ †
Article Snippet: For amplification of hantavirus genes, a two-step PCR was performed in 20-μl reaction mixtures containing 250 μM deoxynucleoside triphosphate (dNTP), 2 mM MgCl2 , 1 U of AmpliTaq polymerase (Roche, Basel, Switzerland), and a 0.25 μM concentration of each primer. .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 s, 2°C step-down annealing from 48°C to 38°C for 40 s, and elongation at 72°C for 1 min and then 32 cycles of denaturation at 94°C for 40 s, annealing at 42°C for 40 s, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Article Title: Molecular Phylogenetic Analysis of Orientia tsutsugamushi Based on the groES and groEL Genes
Article Snippet: In addition to the 56-kD TSA gene, the groES and groEL genes were also amplified. .. Initial denaturation was conducted at 94°C for 2 min, followed by 30 cycles of denaturation at 94°C for 40 s, annealing at 55°C for 30 s, and extension at 72°C for 2 min using a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Article Title: Evolutionary Insights from a Genetically Divergent Hantavirus Harbored by the European Common Mole (Talpa europaea)
Article Snippet: After the initial detection of hantavirus RNAs, amplification of the full-length S- and L-genomic segments was accomplished using oligonucleotide primers based on newly acquired sequences. .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

DNA Sequencing:

Article Title: Complete genome sequence and molecular phylogeny of a newfound hantavirus harbored by the Doucet’s musk shrew (Crocidura douceti) in Guinea
Article Snippet: Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany), and DNA sequencing was performed using an ABI Prism 377XL Genetic Analyzer (Applied Biosystems, Foster City, CA).

Article Title: Molecular evolution of Azagny virus, a newfound hantavirus harbored by the West African pygmy shrew (Crocidura obscurior) in C?te d'Ivoire
Article Snippet: Paragraph title: RT-PCR and DNA sequencing ... Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Article Title: Divergent lineage of a novel hantavirus in the banana pipistrelle (Neoromicia nanus) in C?te d'Ivoire
Article Snippet: Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 s, two-degree step-down annealing from 48°C to 38°C for 40 s, and elongation at 72°C for 1 min or 1 min 20 s, then 32 cycles of denaturation at 94°C for 40 s, annealing at 42°C for 40 s, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany), and DNA sequencing was performed using an ABI Prism 377XL Genetic Analyzer (Applied Biosystems, Foster City, CA).

Article Title: Evolutionary Insights from a Genetically Divergent Hantavirus Harbored by the European Common Mole (Talpa europaea)
Article Snippet: Paragraph title: RT-PCR and DNA Sequencing ... Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

DNA Purification:

Article Title: Genetic basis for retention of a critical virulence plasmid of Borrelia burgdorferi
Article Snippet: The PCR conditions were 94°C for 2 min, followed by 30 cycles of 94°C for 45 s, 55°C for 45 s and 68°C for 3 min in a GeneAmp PCR 9700 thermal cycler (Perkin Elmer) or a DNA engine tetrad 2 thermal cycler (MJ Research). .. Total genomic DNA was then isolated from these cultures with a Wizard genomic DNA purification kit (Promega).

Gel Extraction:

Article Title: Complete genome sequence and molecular phylogeny of a newfound hantavirus harbored by the Doucet’s musk shrew (Crocidura douceti) in Guinea
Article Snippet: Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany), and DNA sequencing was performed using an ABI Prism 377XL Genetic Analyzer (Applied Biosystems, Foster City, CA).

Article Title: Molecular evolution of Azagny virus, a newfound hantavirus harbored by the West African pygmy shrew (Crocidura obscurior) in C?te d'Ivoire
Article Snippet: Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany), and DNA sequencing was performed using an ABI Prism 377XL Genetic Analyzer (Applied Biosystems, Foster City, CA).

Article Title: Complex History of Codiversification and Host Switching of a Newfound Soricid-Borne Orthohantavirus in North America
Article Snippet: Initial denaturation was followed by touchdown cycling (two-degree step-down annealing from 48 °C to 38 °C for 40 sec) and elongation at 72 °C for 1 min, then 32 cycles of denaturation at 94 °C for 40 sec, annealing at 42 °C for 40 sec, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA, USA). .. Amplified products were separated by electrophoresis on 1.5% agarose gels and purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany).

Article Title: Divergent lineage of a novel hantavirus in the banana pipistrelle (Neoromicia nanus) in C?te d'Ivoire
Article Snippet: Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 s, two-degree step-down annealing from 48°C to 38°C for 40 s, and elongation at 72°C for 1 min or 1 min 20 s, then 32 cycles of denaturation at 94°C for 40 s, annealing at 42°C for 40 s, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany), and DNA sequencing was performed using an ABI Prism 377XL Genetic Analyzer (Applied Biosystems, Foster City, CA).

Article Title: Shared Ancestry between a Newfound Mole-Borne Hantavirus and Hantaviruses Harbored by Cricetid Rodents ▿Shared Ancestry between a Newfound Mole-Borne Hantavirus and Hantaviruses Harbored by Cricetid Rodents ▿ †
Article Snippet: Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 s, 2°C step-down annealing from 48°C to 38°C for 40 s, and elongation at 72°C for 1 min and then 32 cycles of denaturation at 94°C for 40 s, annealing at 42°C for 40 s, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were separated by electrophoresis on 1.5% agarose gels and purified using the QIAquick gel extraction kit (Qiagen, Hilden, Germany).

Article Title: Evolutionary Insights from a Genetically Divergent Hantavirus Harbored by the European Common Mole (Talpa europaea)
Article Snippet: Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were separated by electrophoresis on 1.5% agarose gels and purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany).

Reverse Transcription Polymerase Chain Reaction:

Article Title: Divergent ancestral lineages of newfound hantaviruses harbored by phylogenetically related crocidurine shrew species in Korea
Article Snippet: Paragraph title: RNA extraction, cDNA synthesis and RT-PCR amplification ... Initial denaturation at 94°C for 2 min was followed by two cycles each of denaturation at 94°C for 30 sec, two-degree step-down annealing from 46°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 30 cycles of denaturation at 94°C for 30 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA) ( ).

Article Title: Genetic variants of Cao Bang hantavirus in the Chinese mole shrew (Anourosorex squamipes) and Taiwanese mole shrew (Anourosorex yamashinai)
Article Snippet: Total RNA was extracted from tissues using the PureLink Micro-to-Midi total RNA purification kit (Invitrogen, San Diego, CA), and cDNA was synthesized using the SuperScript III First-Strand Synthesis Systems (Invitrogen) with a highly conserved primer and/or random hexamers by two-step RT-PCR cycles. .. Initial denaturation at 94°C for 2 min was followed by two cycles each of denaturation at 94°C for 30 sec, two-degree step-down annealing from 46°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 30 cycles of denaturation at 94°C for 30 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Article Title: Molecular evolution of Azagny virus, a newfound hantavirus harbored by the West African pygmy shrew (Crocidura obscurior) in C?te d'Ivoire
Article Snippet: Paragraph title: RT-PCR and DNA sequencing ... Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Article Title: Complex History of Codiversification and Host Switching of a Newfound Soricid-Borne Orthohantavirus in North America
Article Snippet: Paragraph title: 2.2. RNA Extraction, cDNA Synthesis, and RT-PCR Amplification ... Initial denaturation was followed by touchdown cycling (two-degree step-down annealing from 48 °C to 38 °C for 40 sec) and elongation at 72 °C for 1 min, then 32 cycles of denaturation at 94 °C for 40 sec, annealing at 42 °C for 40 sec, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA, USA).

Article Title: Divergent lineage of a novel hantavirus in the banana pipistrelle (Neoromicia nanus) in C?te d'Ivoire
Article Snippet: Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 s, two-degree step-down annealing from 48°C to 38°C for 40 s, and elongation at 72°C for 1 min or 1 min 20 s, then 32 cycles of denaturation at 94°C for 40 s, annealing at 42°C for 40 s, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. After innumerable failed attempts, hantavirus RNA was detected by RT-PCR in ethanol-fixed liver tissues from two of 12 banana pipistrelles (Neoromicia nanus Peters 1852), captured during June 2011 near Mouyassué village (5°22'07"N, 3°05'37"W) in Aboisso District, 130 km from Abidjan, in the extreme southeastern region of Côte d'Ivoire in West Africa (Figure ).

Article Title: Highly Divergent Genetic Variants of Soricid-Borne Altai Virus (Hantaviridae) in Eurasia Suggest Ancient Host-Switching Events
Article Snippet: Paragraph title: 2.2. RNA Extraction and RT-PCR Analysis ... Initial denaturation at 94 °C for 5 min was followed by two cycles each of denaturation at 94 °C for 40 s, two-degree step-down annealing from 48 to 38 °C for 40 s, and elongation at 72 °C for 1 min, then 32 cycles of denaturation at 94 °C for 40 s, annealing at 42 °C for 40 s, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA, USA).

Article Title: Dahonggou Creek virus, a divergent lineage of hantavirus harbored by the long-tailed mole (Scaptonyx fusicaudus)
Article Snippet: Briefly, total RNA was extracted from tissues using the PureLink Micro-to-Midi total RNA purification kit (Invitrogen, San Diego, CA), and cDNA was synthesized using the SuperScript III First-Strand Synthesis Systems (Invitrogen) with a highly conserved primer and/or random hexamers by two-step RT-PCR cycles. .. Initial denaturation at 94 °C for 2 min was followed by two cycles each of denaturation at 94 °C for 30 s, two-degree step-down annealing from 46 to 38 °C for 40 s, and elongation at 72 °C for 1 min, then 30 cycles of denaturation at 94 °C for 30 s, annealing at 42 °C for 40 s, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (PerkinElmer, Waltham, MA).

Article Title: Novel Hantavirus in the Flat-Skulled Shrew (Sorex roboratus)
Article Snippet: In an attempt to corroborate decades-old reports of hantaviral antigens in shrews from Russia (Gavrilovskaya et al. , Tkachenko et al. , Miasnikov et al. ), archival liver or lung tissues from 4 Siberian large-toothed shrews ( Sorex daphaenodon ), 5 Eurasian least shrews ( Sorex minutissimus ), 12 flat-skulled shrews ( Sorex roboratus ), and 18 tundra shrews ( Sorex tundrensis ), collected as part of the Beringian Coevolution Project in the Sakha Republic in northeastern Siberia ( ) during July and August 2006, were analyzed for hantavirus RNA by reverse transcription–polymerase chain reaction (PCR), using primers designed from highly conserved regions of hantavirus genomes (Arai et al. , Kang et al. , ). cDNA was synthesized using the SuperScript III First-Strand Synthesis System (Invitrogen, San Diego, CA) and an oligonucleotide primer (OSM55, 5′-TAGTAGTAGACTCC-3′) designed from the conserved 3′-end of the S, M, and L segments of hantaviruses. .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 s, two-degree step-down annealing from 48°C to 38°C for 40 s, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 s, annealing at 42°C for 40 s, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Article Title: Shared Ancestry between a Newfound Mole-Borne Hantavirus and Hantaviruses Harbored by Cricetid Rodents ▿Shared Ancestry between a Newfound Mole-Borne Hantavirus and Hantaviruses Harbored by Cricetid Rodents ▿ †
Article Snippet: Paragraph title: RNA extraction and reverse transcription (RT)-PCR analysis. ... Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 s, 2°C step-down annealing from 48°C to 38°C for 40 s, and elongation at 72°C for 1 min and then 32 cycles of denaturation at 94°C for 40 s, annealing at 42°C for 40 s, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Article Title: Evolutionary Insights from a Genetically Divergent Hantavirus Harbored by the European Common Mole (Talpa europaea)
Article Snippet: Paragraph title: RT-PCR and DNA Sequencing ... Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Staining:

Article Title: Genetic basis for retention of a critical virulence plasmid of Borrelia burgdorferi
Article Snippet: The PCR conditions were 94°C for 2 min, followed by 30 cycles of 94°C for 45 s, 55°C for 45 s and 68°C for 3 min in a GeneAmp PCR 9700 thermal cycler (Perkin Elmer) or a DNA engine tetrad 2 thermal cycler (MJ Research). .. PCR products were separated by agarose gel electrophoresis and visualized by ethidium bromide staining.

Concentration Assay:

Article Title: Shared Ancestry between a Newfound Mole-Borne Hantavirus and Hantaviruses Harbored by Cricetid Rodents ▿Shared Ancestry between a Newfound Mole-Borne Hantavirus and Hantaviruses Harbored by Cricetid Rodents ▿ †
Article Snippet: For amplification of hantavirus genes, a two-step PCR was performed in 20-μl reaction mixtures containing 250 μM deoxynucleoside triphosphate (dNTP), 2 mM MgCl2 , 1 U of AmpliTaq polymerase (Roche, Basel, Switzerland), and a 0.25 μM concentration of each primer. .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 s, 2°C step-down annealing from 48°C to 38°C for 40 s, and elongation at 72°C for 1 min and then 32 cycles of denaturation at 94°C for 40 s, annealing at 42°C for 40 s, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA).

Polymerase Chain Reaction:

Article Title: Complete genome sequence and molecular phylogeny of a newfound hantavirus harbored by the Doucet’s musk shrew (Crocidura douceti) in Guinea
Article Snippet: .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany), and DNA sequencing was performed using an ABI Prism 377XL Genetic Analyzer (Applied Biosystems, Foster City, CA).

Article Title: Divergent ancestral lineages of newfound hantaviruses harbored by phylogenetically related crocidurine shrew species in Korea
Article Snippet: .. Initial denaturation at 94°C for 2 min was followed by two cycles each of denaturation at 94°C for 30 sec, two-degree step-down annealing from 46°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 30 cycles of denaturation at 94°C for 30 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA) ( ). .. PCR products were separated, using MobiSpin S-400 spin columns (MoBiTec, Goettingen, Germany), and amplicons were sequenced directly using an ABI Prism 3130 Genetic Analyzer (Applied Biosystems, Foster City, CA).

Article Title: Genetic variants of Cao Bang hantavirus in the Chinese mole shrew (Anourosorex squamipes) and Taiwanese mole shrew (Anourosorex yamashinai)
Article Snippet: .. Initial denaturation at 94°C for 2 min was followed by two cycles each of denaturation at 94°C for 30 sec, two-degree step-down annealing from 46°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 30 cycles of denaturation at 94°C for 30 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. PCR products were separated, using MobiSpin S-400 spin columns (MoBiTec, Goettingen, Germany), and amplicons were sequenced directly using an ABI Prism 3130 Genetic Analyzer (Applied Biosystems, Foster City, CA).

Article Title: Molecular evolution of Azagny virus, a newfound hantavirus harbored by the West African pygmy shrew (Crocidura obscurior) in C?te d'Ivoire
Article Snippet: .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany), and DNA sequencing was performed using an ABI Prism 377XL Genetic Analyzer (Applied Biosystems, Foster City, CA).

Article Title: Isolation and partial characterization of a highly divergent lineage of hantavirus from the European mole (Talpa europaea)
Article Snippet: .. Initial denaturation at 94 °C for 2 min was followed by two cycles each of denaturation at 94 °C for 30 sec, two-degree step-down annealing from 46 °C to 38 °C for 40 sec, and elongation at 72 °C for 1 min, then 30 cycles of denaturation at 94 °C for 30 sec, annealing at 42 °C for 40 sec, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA) . .. PCR products were separated, using MobiSpin S-400 spin columns (MoBiTec, Goettingen, Germany), and amplicons were sequenced directly using an ABI Prism 3130 Genetic Analyzer (Applied Biosystems, Foster City, CA) .

Article Title: Genetic basis for retention of a critical virulence plasmid of Borrelia burgdorferi
Article Snippet: .. The PCR conditions were 94°C for 2 min, followed by 30 cycles of 94°C for 45 s, 55°C for 45 s and 68°C for 3 min in a GeneAmp PCR 9700 thermal cycler (Perkin Elmer) or a DNA engine tetrad 2 thermal cycler (MJ Research). .. PCR products were separated by agarose gel electrophoresis and visualized by ethidium bromide staining.

Article Title: Complex History of Codiversification and Host Switching of a Newfound Soricid-Borne Orthohantavirus in North America
Article Snippet: .. Initial denaturation was followed by touchdown cycling (two-degree step-down annealing from 48 °C to 38 °C for 40 sec) and elongation at 72 °C for 1 min, then 32 cycles of denaturation at 94 °C for 40 sec, annealing at 42 °C for 40 sec, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA, USA). .. Amplified products were separated by electrophoresis on 1.5% agarose gels and purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany).

Article Title: Divergent lineage of a novel hantavirus in the banana pipistrelle (Neoromicia nanus) in C?te d'Ivoire
Article Snippet: .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 s, two-degree step-down annealing from 48°C to 38°C for 40 s, and elongation at 72°C for 1 min or 1 min 20 s, then 32 cycles of denaturation at 94°C for 40 s, annealing at 42°C for 40 s, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany), and DNA sequencing was performed using an ABI Prism 377XL Genetic Analyzer (Applied Biosystems, Foster City, CA).

Article Title: Dahonggou Creek virus, a divergent lineage of hantavirus harbored by the long-tailed mole (Scaptonyx fusicaudus)
Article Snippet: .. Initial denaturation at 94 °C for 2 min was followed by two cycles each of denaturation at 94 °C for 30 s, two-degree step-down annealing from 46 to 38 °C for 40 s, and elongation at 72 °C for 1 min, then 30 cycles of denaturation at 94 °C for 30 s, annealing at 42 °C for 40 s, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (PerkinElmer, Waltham, MA). .. PCR products were separated, using MobiSpin S-400 spin columns (MoBiTec, Goettingen, Germany), and amplicons were sequenced directly using an ABI Prism 3130 Genetic Analyzer (Applied Biosystems, Foster City, CA).

Article Title: Molecular Phylogeny of Hantaviruses Harbored by Insectivorous Bats in C?te d'Ivoire and Vietnam
Article Snippet: .. Initial denaturation at 94 °C for 2 min was followed by two cycles each of denaturation at 94 °C for 30 s, two-degree step-down annealing from 46 °C to 38 °C for 40 s, and elongation at 72 °C for 1 min, then 30 cycles of denaturation at 94 °C for 30 s, annealing at 42 °C for 40 s, and elongation at 72 °C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA, USA) [ , , , , ]. .. PCR products, separated using MobiSpin S-400 spin columns (MoBiTec, Goettingen, Germany), were sequenced directly using an ABI Prism 3130 Genetic Analyzer (Applied Biosystems, Foster City, CA, USA) [ , ].

Article Title: Shared Ancestry between a Newfound Mole-Borne Hantavirus and Hantaviruses Harbored by Cricetid Rodents ▿Shared Ancestry between a Newfound Mole-Borne Hantavirus and Hantaviruses Harbored by Cricetid Rodents ▿ †
Article Snippet: .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 s, 2°C step-down annealing from 48°C to 38°C for 40 s, and elongation at 72°C for 1 min and then 32 cycles of denaturation at 94°C for 40 s, annealing at 42°C for 40 s, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were separated by electrophoresis on 1.5% agarose gels and purified using the QIAquick gel extraction kit (Qiagen, Hilden, Germany).

Article Title: Molecular Phylogenetic Analysis of Orientia tsutsugamushi Based on the groES and groEL Genes
Article Snippet: .. Initial denaturation was conducted at 94°C for 2 min, followed by 30 cycles of denaturation at 94°C for 40 s, annealing at 55°C for 30 s, and extension at 72°C for 2 min using a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. PCR products were separated with MobiSpin S-400 spin columns (MoBiTec, Goettingen, Germany), and amplicons were sequenced directly using an ABI Prism 3130 Genetic Analyzer (Applied Biosystems, Foster City, CA).

Article Title: Evolutionary Insights from a Genetically Divergent Hantavirus Harbored by the European Common Mole (Talpa europaea)
Article Snippet: .. Initial denaturation at 94°C for 5 min was followed by two cycles each of denaturation at 94°C for 40 sec, two-degree step-down annealing from 48°C to 38°C for 40 sec, and elongation at 72°C for 1 min, then 32 cycles of denaturation at 94°C for 40 sec, annealing at 42°C for 40 sec, and elongation at 72°C for 1 min, in a GeneAmp PCR 9700 thermal cycler (Perkin-Elmer, Waltham, MA). .. Amplicons were separated by electrophoresis on 1.5% agarose gels and purified using the QIAQuick Gel Extraction Kit (Qiagen, Hilden, Germany).

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