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Dntp, supplied by Bio Basic Canada, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Clone Assay:

Article Title: Exhausting exercise and tissue-specific expression of monocarboxylate transporters in rainbow trout
Article Snippet: Paragraph title: Cloning, sequencing, and partial characterization of MCTs. ... After cDNA synthesis, a PCR reaction was performed as follows: 1× PCR buffer, 1.5 mM MgSO4 , 0.2 mM dNTP, 0.2 mM of Zfish-MCT F and Zfish-MCT R each, 2 μl cDNA, 2 U Taq polymerase (Bio Basic; Amherst, NY), and DNase/RNase-free water for a final volume of 50 μl.

Article Title: Cis-acting requirements in flanking DNA for the programmed elimination of mse2.9: a common mechanism for deletion of internal eliminated sequences from the developing macronucleus of Tetrahymena thermophila
Article Snippet: The primers 5r and RU4 were used to amplify processed mse2.9 junction sequences in transformed strains, except in the deletion clones where the primers Lba2 and 3r were used. .. PCR was performed in a 50 µl vol containing 0.5 µM each primer, 0.2 mM each dNTP, 10× tsg buffer, 1.5 mM MgCl2 and 2.5 U tsg (Biobasic, Toronto, Canada).

Article Title: Exhausting exercise and tissue-specific expression of monocarboxylate transporters in rainbow trout
Article Snippet: .. Clones (24 from brain and 28 from white muscle) were picked and screened by PCR in a reaction mix containing: 1× PCR buffer, 1.5 mM MgSO4 , 0.2 mM dNTP, 0.2 mM of M13 forward and reverse primers, 2 U Taq polymerase (Bio Basic), and DNase/RNase-free water to a final volume of 50 μl. .. The thermal profile was as follows: an incubation step (94°C/5 min), followed by 40 cycles (94°C/30 s; 55°C/30 s; 72°C/1 min), and a final extension (72°C/5 min).

Centrifugation:

Article Title: Calorie restriction effects on circadian rhythms in gene expression are sex dependent
Article Snippet: Following chloroform extraction step, total RNA was precipitated with isopropanol by centrifugation. .. The RT mix was prepared using 1 µg of mRNA, 50 ng of random hexamer (N808127, Invitrogen), 10 mM dNTP (DD0058, Biobasic), 0.1 M DDT and RNaseOUT Recombinant RNase inhibitor (10777-019, 40 units/ µl) and it was reverse transcribed using SuperScript III Reverse Transcriptase (18080-044, Invitrogen).

Amplification:

Article Title: Multiple Mating and Family Structure of the Western Tent Caterpillar, Malacosoma californicum pluviale: Impact on Disease Resistance
Article Snippet: PCR reactions used 10 ng of genomic DNA, 0.0625 mM dNTP, (1X) PCR buffer (Bio Basic Inc. ON, Canada), 0.25 units of Taq (Bio Basic Inc.), 2 mM MgCl2 , 3 pmol forward and reverse primers (Eurofins MGW Operon, AL, USA), 6.25 μL H2 0 for a total reaction volume of 10 μL. .. PCR conditions were as follows: 95°C for 3 min, cycled 35 times at 94°C for 40 sec, 56°C for 40 sec, 72°C for 30 sec, with a final extension of 72°C for 4 min. Four, 10 μL reactions were performed for each individual sequenced and amplified products were pooled prior to purification with a sodium acetate precipitation, and sent to Eurofins MWG Operon (AL, USA) for sequencing.

Article Title: The origin and population genetic structure of the ‘golden tide’ seaweeds, Sargassum horneri, in Korean waters
Article Snippet: .. Polymerase chain reaction (PCR) amplification was carried out in a reaction volume of 15 μl containing 1 × PCR buffer, 25 μM of each dNTP (Bio Basic Inc., Markham, ON, Canada), 0.6 μM of each of the forward and reverse primers, 0.2 units of Taq DNA polymerase (Thermo Fisher Scientific, Waltham, MA, USA) and approximately 5−10 ng of genomic DNA. .. RCR thermal cycling conditions comprised an initial denaturation at 94 °C for 3 min, followed by 35 cycles of denaturation at 94 °C for 20 s, annealing at 50 °C for 50 s and extension at 68 °C for 60 s, and a final extension at 68 °C for 10 min. PCR products were checked on 2% agarose gels stained with Redsafe (iNtRON).

Article Title: Population genetic structure of eelgrass (Zostera marina) on the Korean coast: Current status and conservation implications for future management
Article Snippet: .. PCR amplification was accomplished in a reaction volume of 15 μl containing 25 μM of each dNTP (Bio Basic), 0.6 μM each of the forward and reverse primers, 0.2 units of Taq DNA polymerase (Thermo Fisher Scientific), 1× PCR buffer, and approximately 5−10 ng of template DNA. .. PCR cycling conditions comprised an initial denaturation phase at 94°C for 5 min, followed by 37 cycles of 94°C for 20 sec (denaturation), 54−57°C for 30 sec (annealing), and 72°C for 30 sec (extension), followed by a terminal extension phase at 72°C for 12 min in a 2720 thermal cycler (Applied Biosystems).

Article Title: The role of structural pleiotropy and regulatory evolution in the retention of heteromers of paralogs
Article Snippet: Tubes were centrifuged for 5 min at 1800 g and 2.5 µL of supernatant was added to a PCR mix composed of 16.85 µL of DNAse free water, 2.5 µL of 10X Taq buffer (BioShop Canada Inc, Canada), 1.5 µL of 25 mM MgCl2, 0.5 µL of 10 mM dNTP (Bio Basic Inc, Canada), 0.15 µL of 5 U/µL Taq DNA polymerase (BioShop Canada Inc, Canada), 0.5 µL of 10 µM Oligo-C and 0.5 µL of 10 µM ADHterm_R. .. The DHFR fragments and associated resistance modules were amplified from plasmids pAG25-linker-F[1,2]-ADHterm (NAT resistance marker) and pAG32-linker-F[3]-ADHterm (HygB resistance marker) ( ) using oligonucleotides defined in ( Table S12).

Article Title: Contrasting life histories contribute to divergent patterns of genetic diversity and population connectivity in freshwater sculpin fishes
Article Snippet: .. Polymerase chain reaction (PCR) amplification was performed in a reaction volume of 15 μl comprising 1× PCR buffer, 25 μM of each dNTP (Bio Basic Inc., Markham, ON, Canada), 0.6 μM of each of the forward and reverse primers and 0.2 U of Taq polymerase (Thermo Fisher Scientific, Waltham, MA, USA). .. The following thermal cycling conditions were used: initial denaturation at 94 °C for 1 min followed by 35 cycles of denaturation at 94 °C for 1 min, annealing at 58 °C for 1 min and extension at 72 °C for 1 min, followed by a final extension at 72 °C for 20 min in a 2720 thermal cycler (Applied Biosystems, Foster City, CA, USA).

Article Title: Multiple Mating and Family Structure of the Western Tent Caterpillar, Malacosoma californicum pluviale: Impact on Disease Resistance
Article Snippet: PCR reactions for all microsatellites used 10 ng of genomic DNA, 0.0625 mM dNTP, (1X) PCR Reaction buffer (Bio Basic Inc. ON, Canada), 0.25 units of Taq (Bio Basics Inc.), 0.75 or 1 mM MgCl2 (Fermentas, ON, Canada; optimized for each locus, see ), 0.15 pmol of fluorescently labeled forward primer (IRDye® 700 or 800, Integrated DNA Technologies, CA, USA), 3 pmol reverse primer (Eurofins MGW Operon, Huntsville, AL, USA), 6.15 or 6.25 μL H2 0 for a total reaction volume of 10 μL. .. Amplified products were denatured and run on a 6% polyacrylamide gel electrophoresis for 1.5 hours on a LI-COR 4300 automated sequencer (LICOR Inc., NE, USA) with a minimum of four size standards (50–350 bp or 50–700 bp LICOR Inc.) per 64 well gel and two to four reference samples.

Random Hexamer Labeling:

Article Title: Calorie restriction effects on circadian rhythms in gene expression are sex dependent
Article Snippet: .. The RT mix was prepared using 1 µg of mRNA, 50 ng of random hexamer (N808127, Invitrogen), 10 mM dNTP (DD0058, Biobasic), 0.1 M DDT and RNaseOUT Recombinant RNase inhibitor (10777-019, 40 units/ µl) and it was reverse transcribed using SuperScript III Reverse Transcriptase (18080-044, Invitrogen). .. RNA quantification was performed using qPCR with Universal SYBR Green Mix (1725125, BioRad).

Quantitative RT-PCR:

Article Title: Tissue-Specific Localization NUCB2/nesfatin-1 in the Liver and Heart of Mouse Fetus
Article Snippet: .. First strand cDNA synthesis was performed using 2 μg RNA, 10 pmol oligo dT and RNase-free DEPC solution at 70° for 5min, followed by double-strand synthesis in 5× RT buffer (Invitrogen, Carlsbad, CA, USA) with 8 mM dNTP (BIO BASIC INC., Ontario, Canada), 200 unit/μL RTase (Invitrogen, Carlsbad, CA, USA) and RNase-free DEPC solution at 37° for 60 min and at 72° for 15 min. qRT-PCR was performed in a total volume of 20 μL buffer solution containing 2 μL of template cDNA, 10 μL of SYBR Green (Roche, Manheim, Germany), and 10 pmol of each primer. ..

Real-time Polymerase Chain Reaction:

Article Title: Functional Evaluation of Genetic and Environmental Regulators of P450 mRNA Levels
Article Snippet: Paragraph title: Absolute Quantitation of mRNA level by Real-time PCR ... Reverse transcription reactions were prepared in a total volume of 20 µl containing 1× RT buffer, 0.5 mM dNTP, 0.5 µM poly24dT, 5 µM N9, 20 U RNase inhibitor (BIO BASIC INC.) and 100 U M-MLV(H-) reverse transcriptase (Promega).

Article Title: Tissue-Specific Localization NUCB2/nesfatin-1 in the Liver and Heart of Mouse Fetus
Article Snippet: First strand cDNA synthesis was performed using 2 μg RNA, 10 pmol oligo dT and RNase-free DEPC solution at 70° for 5min, followed by double-strand synthesis in 5× RT buffer (Invitrogen, Carlsbad, CA, USA) with 8 mM dNTP (BIO BASIC INC., Ontario, Canada), 200 unit/μL RTase (Invitrogen, Carlsbad, CA, USA) and RNase-free DEPC solution at 37° for 60 min and at 72° for 15 min. qRT-PCR was performed in a total volume of 20 μL buffer solution containing 2 μL of template cDNA, 10 μL of SYBR Green (Roche, Manheim, Germany), and 10 pmol of each primer. .. The optimal temperature cycling protocol was determined to be 95° for 5 min followed by 45 reaction cycles at 95° for 10 s, 60° for 10 s and 72° for 10 s using the LightCycler® 480 Real-time PCR System (Roche, Manheim, Germany).

Article Title: Calorie restriction effects on circadian rhythms in gene expression are sex dependent
Article Snippet: The RT mix was prepared using 1 µg of mRNA, 50 ng of random hexamer (N808127, Invitrogen), 10 mM dNTP (DD0058, Biobasic), 0.1 M DDT and RNaseOUT Recombinant RNase inhibitor (10777-019, 40 units/ µl) and it was reverse transcribed using SuperScript III Reverse Transcriptase (18080-044, Invitrogen). .. RNA quantification was performed using qPCR with Universal SYBR Green Mix (1725125, BioRad).

Incubation:

Article Title: Exhausting exercise and tissue-specific expression of monocarboxylate transporters in rainbow trout
Article Snippet: Clones (24 from brain and 28 from white muscle) were picked and screened by PCR in a reaction mix containing: 1× PCR buffer, 1.5 mM MgSO4 , 0.2 mM dNTP, 0.2 mM of M13 forward and reverse primers, 2 U Taq polymerase (Bio Basic), and DNase/RNase-free water to a final volume of 50 μl. .. The thermal profile was as follows: an incubation step (94°C/5 min), followed by 40 cycles (94°C/30 s; 55°C/30 s; 72°C/1 min), and a final extension (72°C/5 min).

Expressing:

Article Title: Exhausting exercise and tissue-specific expression of monocarboxylate transporters in rainbow trout
Article Snippet: Therefore, total RNA (from two fish not used in the gene expression experiments) was isolated from white muscle and brain using the TRIzol method (Invitrogen; Carlsbad, CA). .. After cDNA synthesis, a PCR reaction was performed as follows: 1× PCR buffer, 1.5 mM MgSO4 , 0.2 mM dNTP, 0.2 mM of Zfish-MCT F and Zfish-MCT R each, 2 μl cDNA, 2 U Taq polymerase (Bio Basic; Amherst, NY), and DNase/RNase-free water for a final volume of 50 μl.

Article Title: Calorie restriction effects on circadian rhythms in gene expression are sex dependent
Article Snippet: Paragraph title: RNA isolation and analysis of mRNA expression ... The RT mix was prepared using 1 µg of mRNA, 50 ng of random hexamer (N808127, Invitrogen), 10 mM dNTP (DD0058, Biobasic), 0.1 M DDT and RNaseOUT Recombinant RNase inhibitor (10777-019, 40 units/ µl) and it was reverse transcribed using SuperScript III Reverse Transcriptase (18080-044, Invitrogen).

Transformation Assay:

Article Title: Exhausting exercise and tissue-specific expression of monocarboxylate transporters in rainbow trout
Article Snippet: After cDNA synthesis, a PCR reaction was performed as follows: 1× PCR buffer, 1.5 mM MgSO4 , 0.2 mM dNTP, 0.2 mM of Zfish-MCT F and Zfish-MCT R each, 2 μl cDNA, 2 U Taq polymerase (Bio Basic; Amherst, NY), and DNase/RNase-free water for a final volume of 50 μl. .. The amplicons [565 base pair (bp)] obtained from both brain and white muscle were subcloned into TopoTA vector (Invitrogen) and transformed in chemically competent Escherichia coli cells.

Article Title: Cis-acting requirements in flanking DNA for the programmed elimination of mse2.9: a common mechanism for deletion of internal eliminated sequences from the developing macronucleus of Tetrahymena thermophila
Article Snippet: These primer sets amplify junctions only in transformed strains, as 3r and 5r are complementary to rDNA sequence. .. PCR was performed in a 50 µl vol containing 0.5 µM each primer, 0.2 mM each dNTP, 10× tsg buffer, 1.5 mM MgCl2 and 2.5 U tsg (Biobasic, Toronto, Canada).

DNA Sequencing:

Article Title: The origin and population genetic structure of the ‘golden tide’ seaweeds, Sargassum horneri, in Korean waters
Article Snippet: Paragraph title: Mitochondrial (mt) DNA sequencing ... Polymerase chain reaction (PCR) amplification was carried out in a reaction volume of 15 μl containing 1 × PCR buffer, 25 μM of each dNTP (Bio Basic Inc., Markham, ON, Canada), 0.6 μM of each of the forward and reverse primers, 0.2 units of Taq DNA polymerase (Thermo Fisher Scientific, Waltham, MA, USA) and approximately 5−10 ng of genomic DNA.

Polymerase Chain Reaction:

Article Title: Multiple Mating and Family Structure of the Western Tent Caterpillar, Malacosoma californicum pluviale: Impact on Disease Resistance
Article Snippet: .. PCR reactions used 10 ng of genomic DNA, 0.0625 mM dNTP, (1X) PCR buffer (Bio Basic Inc. ON, Canada), 0.25 units of Taq (Bio Basic Inc.), 2 mM MgCl2 , 3 pmol forward and reverse primers (Eurofins MGW Operon, AL, USA), 6.25 μL H2 0 for a total reaction volume of 10 μL. .. PCR conditions were as follows: 95°C for 3 min, cycled 35 times at 94°C for 40 sec, 56°C for 40 sec, 72°C for 30 sec, with a final extension of 72°C for 4 min. Four, 10 μL reactions were performed for each individual sequenced and amplified products were pooled prior to purification with a sodium acetate precipitation, and sent to Eurofins MWG Operon (AL, USA) for sequencing.

Article Title: The origin and population genetic structure of the ‘golden tide’ seaweeds, Sargassum horneri, in Korean waters
Article Snippet: .. Polymerase chain reaction (PCR) amplification was carried out in a reaction volume of 15 μl containing 1 × PCR buffer, 25 μM of each dNTP (Bio Basic Inc., Markham, ON, Canada), 0.6 μM of each of the forward and reverse primers, 0.2 units of Taq DNA polymerase (Thermo Fisher Scientific, Waltham, MA, USA) and approximately 5−10 ng of genomic DNA. .. RCR thermal cycling conditions comprised an initial denaturation at 94 °C for 3 min, followed by 35 cycles of denaturation at 94 °C for 20 s, annealing at 50 °C for 50 s and extension at 68 °C for 60 s, and a final extension at 68 °C for 10 min. PCR products were checked on 2% agarose gels stained with Redsafe (iNtRON).

Article Title: Exhausting exercise and tissue-specific expression of monocarboxylate transporters in rainbow trout
Article Snippet: .. After cDNA synthesis, a PCR reaction was performed as follows: 1× PCR buffer, 1.5 mM MgSO4 , 0.2 mM dNTP, 0.2 mM of Zfish-MCT F and Zfish-MCT R each, 2 μl cDNA, 2 U Taq polymerase (Bio Basic; Amherst, NY), and DNase/RNase-free water for a final volume of 50 μl. .. The thermal profile was started with 2 cycles (94°C/2 min; 63°C/1 min; 72°C/1 min), followed by 35 cycles (94°C/30 s; 63°C/30 s; 72°C/1 min), and a final extension step (72°C/5 min) using either Eppendorf Mastercycler (Hamburg, Germany) or Bio-Rad S1000 thermal cycler (Hercules, CA).

Article Title: Cis-acting requirements in flanking DNA for the programmed elimination of mse2.9: a common mechanism for deletion of internal eliminated sequences from the developing macronucleus of Tetrahymena thermophila
Article Snippet: .. PCR was performed in a 50 µl vol containing 0.5 µM each primer, 0.2 mM each dNTP, 10× tsg buffer, 1.5 mM MgCl2 and 2.5 U tsg (Biobasic, Toronto, Canada). .. PCR was performed in a Perkin-Elmer 9600 thermal cycler using the following cycling conditions: one cycle at 94°C for 3 min; 35 cycles at 94°C for 30 s, 55°C for 30 s, 72°C for 30 s; one cycle at 72°C for 5 min. Long PCR was performed with the Expand Long Template PCR system (Boehringer Mannheim), using conditions as specified by the manufacturer.

Article Title: Population genetic structure of eelgrass (Zostera marina) on the Korean coast: Current status and conservation implications for future management
Article Snippet: .. PCR amplification was accomplished in a reaction volume of 15 μl containing 25 μM of each dNTP (Bio Basic), 0.6 μM each of the forward and reverse primers, 0.2 units of Taq DNA polymerase (Thermo Fisher Scientific), 1× PCR buffer, and approximately 5−10 ng of template DNA. .. PCR cycling conditions comprised an initial denaturation phase at 94°C for 5 min, followed by 37 cycles of 94°C for 20 sec (denaturation), 54−57°C for 30 sec (annealing), and 72°C for 30 sec (extension), followed by a terminal extension phase at 72°C for 12 min in a 2720 thermal cycler (Applied Biosystems).

Article Title: The role of structural pleiotropy and regulatory evolution in the retention of heteromers of paralogs
Article Snippet: .. Tubes were centrifuged for 5 min at 1800 g and 2.5 µL of supernatant was added to a PCR mix composed of 16.85 µL of DNAse free water, 2.5 µL of 10X Taq buffer (BioShop Canada Inc, Canada), 1.5 µL of 25 mM MgCl2, 0.5 µL of 10 mM dNTP (Bio Basic Inc, Canada), 0.15 µL of 5 U/µL Taq DNA polymerase (BioShop Canada Inc, Canada), 0.5 µL of 10 µM Oligo-C and 0.5 µL of 10 µM ADHterm_R. .. We confirmed by PCR 2025 strains from the DHFR collection and 126 strains out of the 154 from ( Tables S9, S10, and S12).

Article Title: Contrasting life histories contribute to divergent patterns of genetic diversity and population connectivity in freshwater sculpin fishes
Article Snippet: .. Polymerase chain reaction (PCR) amplification was performed in a reaction volume of 15 μl comprising 1× PCR buffer, 25 μM of each dNTP (Bio Basic Inc., Markham, ON, Canada), 0.6 μM of each of the forward and reverse primers and 0.2 U of Taq polymerase (Thermo Fisher Scientific, Waltham, MA, USA). .. The following thermal cycling conditions were used: initial denaturation at 94 °C for 1 min followed by 35 cycles of denaturation at 94 °C for 1 min, annealing at 58 °C for 1 min and extension at 72 °C for 1 min, followed by a final extension at 72 °C for 20 min in a 2720 thermal cycler (Applied Biosystems, Foster City, CA, USA).

Article Title: Multiple Mating and Family Structure of the Western Tent Caterpillar, Malacosoma californicum pluviale: Impact on Disease Resistance
Article Snippet: .. PCR reactions for all microsatellites used 10 ng of genomic DNA, 0.0625 mM dNTP, (1X) PCR Reaction buffer (Bio Basic Inc. ON, Canada), 0.25 units of Taq (Bio Basics Inc.), 0.75 or 1 mM MgCl2 (Fermentas, ON, Canada; optimized for each locus, see ), 0.15 pmol of fluorescently labeled forward primer (IRDye® 700 or 800, Integrated DNA Technologies, CA, USA), 3 pmol reverse primer (Eurofins MGW Operon, Huntsville, AL, USA), 6.15 or 6.25 μL H2 0 for a total reaction volume of 10 μL. ..

Article Title: Exhausting exercise and tissue-specific expression of monocarboxylate transporters in rainbow trout
Article Snippet: .. Clones (24 from brain and 28 from white muscle) were picked and screened by PCR in a reaction mix containing: 1× PCR buffer, 1.5 mM MgSO4 , 0.2 mM dNTP, 0.2 mM of M13 forward and reverse primers, 2 U Taq polymerase (Bio Basic), and DNase/RNase-free water to a final volume of 50 μl. .. The thermal profile was as follows: an incubation step (94°C/5 min), followed by 40 cycles (94°C/30 s; 55°C/30 s; 72°C/1 min), and a final extension (72°C/5 min).

Article Title: Environmental and genetic determinants of transcriptional plasticity in Chinook salmon
Article Snippet: .. PCR reactions consisted of 20 mM Tris-HCl pH 8.75, 10 mM KCl, 10 mM (NH4 )2 SO4 , 0.1% Triton X-100, 0.1 mg/mL BSA, 200 μM each dNTP, 200 nM forward and reverse primers, 2.0 mM MgSO4 , 0.5 U of taq polymerase (Bio Basic Canada Inc., Markham, ON) and ~50 ng of DNA. .. Conditions for thermal cycling were 95 °C for 2 min, 35 cycles of 95 °C for 15 s, locus-specific annealing temperature (52 °C–OtsG68, 56 °C–OtsG432, 58 °C–Ots208, 60 °C–Ots209, Ots211) for 15 s and 72 °C for 30 s, followed by 72 °C for 5 min. Microsatellite PCR products were characterized using a Licor 4300 DNA Analyzer (Licor Biosciences Inc.) and Gene ImagR software (Scanalytics Inc.).

Recombinant:

Article Title: Calorie restriction effects on circadian rhythms in gene expression are sex dependent
Article Snippet: .. The RT mix was prepared using 1 µg of mRNA, 50 ng of random hexamer (N808127, Invitrogen), 10 mM dNTP (DD0058, Biobasic), 0.1 M DDT and RNaseOUT Recombinant RNase inhibitor (10777-019, 40 units/ µl) and it was reverse transcribed using SuperScript III Reverse Transcriptase (18080-044, Invitrogen). .. RNA quantification was performed using qPCR with Universal SYBR Green Mix (1725125, BioRad).

DNA Extraction:

Article Title: The origin and population genetic structure of the ‘golden tide’ seaweeds, Sargassum horneri, in Korean waters
Article Snippet: Mitochondrial (mt) DNA sequencing Genomic DNA was extracted from pulverized leaf samples using i-genomic Plant DNA Extraction Mini Kit (iNtRON Biotechnology, Daejeon, Korea). .. Polymerase chain reaction (PCR) amplification was carried out in a reaction volume of 15 μl containing 1 × PCR buffer, 25 μM of each dNTP (Bio Basic Inc., Markham, ON, Canada), 0.6 μM of each of the forward and reverse primers, 0.2 units of Taq DNA polymerase (Thermo Fisher Scientific, Waltham, MA, USA) and approximately 5−10 ng of genomic DNA.

Article Title: Population genetic structure of eelgrass (Zostera marina) on the Korean coast: Current status and conservation implications for future management
Article Snippet: Paragraph title: DNA extraction, PCR, and microsatellite genotyping ... PCR amplification was accomplished in a reaction volume of 15 μl containing 25 μM of each dNTP (Bio Basic), 0.6 μM each of the forward and reverse primers, 0.2 units of Taq DNA polymerase (Thermo Fisher Scientific), 1× PCR buffer, and approximately 5−10 ng of template DNA.

Article Title: Multiple Mating and Family Structure of the Western Tent Caterpillar, Malacosoma californicum pluviale: Impact on Disease Resistance
Article Snippet: Paragraph title: DNA extraction and microsatellite analysis ... PCR reactions for all microsatellites used 10 ng of genomic DNA, 0.0625 mM dNTP, (1X) PCR Reaction buffer (Bio Basic Inc. ON, Canada), 0.25 units of Taq (Bio Basics Inc.), 0.75 or 1 mM MgCl2 (Fermentas, ON, Canada; optimized for each locus, see ), 0.15 pmol of fluorescently labeled forward primer (IRDye® 700 or 800, Integrated DNA Technologies, CA, USA), 3 pmol reverse primer (Eurofins MGW Operon, Huntsville, AL, USA), 6.15 or 6.25 μL H2 0 for a total reaction volume of 10 μL.

Isolation:

Article Title: Exhausting exercise and tissue-specific expression of monocarboxylate transporters in rainbow trout
Article Snippet: Therefore, total RNA (from two fish not used in the gene expression experiments) was isolated from white muscle and brain using the TRIzol method (Invitrogen; Carlsbad, CA). .. After cDNA synthesis, a PCR reaction was performed as follows: 1× PCR buffer, 1.5 mM MgSO4 , 0.2 mM dNTP, 0.2 mM of Zfish-MCT F and Zfish-MCT R each, 2 μl cDNA, 2 U Taq polymerase (Bio Basic; Amherst, NY), and DNase/RNase-free water for a final volume of 50 μl.

Article Title: Th 17 Cells and Nesfatin-1 are associated with Spontaneous Abortion in the CBA/j × DBA/2 Mouse Model
Article Snippet: RNA extraction and cDNA synthesis Total RNA was isolated by using the RNA isoplus (TaKaRa Bio, Shiga, Japan) according to manufacturer’s instruction. .. First strand cDNA synthesis was performed in RNase-free DEPC solution containing 2 μg total RNA and 10 pmol oligo dT at 70°C for 5 min, followed by double-strand synthesis in 5× RT buffer (Invitrogen, Carlsbad, CA) with 8 mM dNTP (BIO BASIC INC., Ontario, Canada), 200 unit/μL RTase (Invitrogen, Carlsbad, CA) at 37°C for 60min and at 72°C for 15 min.

Article Title: Calorie restriction effects on circadian rhythms in gene expression are sex dependent
Article Snippet: Paragraph title: RNA isolation and analysis of mRNA expression ... The RT mix was prepared using 1 µg of mRNA, 50 ng of random hexamer (N808127, Invitrogen), 10 mM dNTP (DD0058, Biobasic), 0.1 M DDT and RNaseOUT Recombinant RNase inhibitor (10777-019, 40 units/ µl) and it was reverse transcribed using SuperScript III Reverse Transcriptase (18080-044, Invitrogen).

Article Title: Contrasting life histories contribute to divergent patterns of genetic diversity and population connectivity in freshwater sculpin fishes
Article Snippet: Genomic DNA was isolated using a DNeasy Blood and Tissue Kit (Qiagen, Hilden, Germany). .. Polymerase chain reaction (PCR) amplification was performed in a reaction volume of 15 μl comprising 1× PCR buffer, 25 μM of each dNTP (Bio Basic Inc., Markham, ON, Canada), 0.6 μM of each of the forward and reverse primers and 0.2 U of Taq polymerase (Thermo Fisher Scientific, Waltham, MA, USA).

Size-exclusion Chromatography:

Article Title: Multiple Mating and Family Structure of the Western Tent Caterpillar, Malacosoma californicum pluviale: Impact on Disease Resistance
Article Snippet: PCR reactions used 10 ng of genomic DNA, 0.0625 mM dNTP, (1X) PCR buffer (Bio Basic Inc. ON, Canada), 0.25 units of Taq (Bio Basic Inc.), 2 mM MgCl2 , 3 pmol forward and reverse primers (Eurofins MGW Operon, AL, USA), 6.25 μL H2 0 for a total reaction volume of 10 μL. .. PCR conditions were as follows: 95°C for 3 min, cycled 35 times at 94°C for 40 sec, 56°C for 40 sec, 72°C for 30 sec, with a final extension of 72°C for 4 min. Four, 10 μL reactions were performed for each individual sequenced and amplified products were pooled prior to purification with a sodium acetate precipitation, and sent to Eurofins MWG Operon (AL, USA) for sequencing.

Article Title: Population genetic structure of eelgrass (Zostera marina) on the Korean coast: Current status and conservation implications for future management
Article Snippet: PCR amplification was accomplished in a reaction volume of 15 μl containing 25 μM of each dNTP (Bio Basic), 0.6 μM each of the forward and reverse primers, 0.2 units of Taq DNA polymerase (Thermo Fisher Scientific), 1× PCR buffer, and approximately 5−10 ng of template DNA. .. PCR cycling conditions comprised an initial denaturation phase at 94°C for 5 min, followed by 37 cycles of 94°C for 20 sec (denaturation), 54−57°C for 30 sec (annealing), and 72°C for 30 sec (extension), followed by a terminal extension phase at 72°C for 12 min in a 2720 thermal cycler (Applied Biosystems).

Labeling:

Article Title: Multiple Mating and Family Structure of the Western Tent Caterpillar, Malacosoma californicum pluviale: Impact on Disease Resistance
Article Snippet: .. PCR reactions for all microsatellites used 10 ng of genomic DNA, 0.0625 mM dNTP, (1X) PCR Reaction buffer (Bio Basic Inc. ON, Canada), 0.25 units of Taq (Bio Basics Inc.), 0.75 or 1 mM MgCl2 (Fermentas, ON, Canada; optimized for each locus, see ), 0.15 pmol of fluorescently labeled forward primer (IRDye® 700 or 800, Integrated DNA Technologies, CA, USA), 3 pmol reverse primer (Eurofins MGW Operon, Huntsville, AL, USA), 6.15 or 6.25 μL H2 0 for a total reaction volume of 10 μL. ..

Purification:

Article Title: Multiple Mating and Family Structure of the Western Tent Caterpillar, Malacosoma californicum pluviale: Impact on Disease Resistance
Article Snippet: PCR reactions used 10 ng of genomic DNA, 0.0625 mM dNTP, (1X) PCR buffer (Bio Basic Inc. ON, Canada), 0.25 units of Taq (Bio Basic Inc.), 2 mM MgCl2 , 3 pmol forward and reverse primers (Eurofins MGW Operon, AL, USA), 6.25 μL H2 0 for a total reaction volume of 10 μL. .. PCR conditions were as follows: 95°C for 3 min, cycled 35 times at 94°C for 40 sec, 56°C for 40 sec, 72°C for 30 sec, with a final extension of 72°C for 4 min. Four, 10 μL reactions were performed for each individual sequenced and amplified products were pooled prior to purification with a sodium acetate precipitation, and sent to Eurofins MWG Operon (AL, USA) for sequencing.

Article Title: The origin and population genetic structure of the ‘golden tide’ seaweeds, Sargassum horneri, in Korean waters
Article Snippet: Polymerase chain reaction (PCR) amplification was carried out in a reaction volume of 15 μl containing 1 × PCR buffer, 25 μM of each dNTP (Bio Basic Inc., Markham, ON, Canada), 0.6 μM of each of the forward and reverse primers, 0.2 units of Taq DNA polymerase (Thermo Fisher Scientific, Waltham, MA, USA) and approximately 5−10 ng of genomic DNA. .. The amplified mtDNA were purified enzymatically with Exonuclease I (New England BioLabs, Ipswich, MA, USA) and Shrimp Alkaline Phosphatase (New England BioLabs).

Article Title: Contrasting life histories contribute to divergent patterns of genetic diversity and population connectivity in freshwater sculpin fishes
Article Snippet: Polymerase chain reaction (PCR) amplification was performed in a reaction volume of 15 μl comprising 1× PCR buffer, 25 μM of each dNTP (Bio Basic Inc., Markham, ON, Canada), 0.6 μM of each of the forward and reverse primers and 0.2 U of Taq polymerase (Thermo Fisher Scientific, Waltham, MA, USA). .. The amplified PCR products were purified enzymatically with Exonuclease I (New England BioLabs, Ipswich, MA, USA) and Shrimp Alkaline Phosphatase (New England BioLabs).

Sequencing:

Article Title: Multiple Mating and Family Structure of the Western Tent Caterpillar, Malacosoma californicum pluviale: Impact on Disease Resistance
Article Snippet: Paragraph title: Sequencing ... PCR reactions used 10 ng of genomic DNA, 0.0625 mM dNTP, (1X) PCR buffer (Bio Basic Inc. ON, Canada), 0.25 units of Taq (Bio Basic Inc.), 2 mM MgCl2 , 3 pmol forward and reverse primers (Eurofins MGW Operon, AL, USA), 6.25 μL H2 0 for a total reaction volume of 10 μL.

Article Title: The origin and population genetic structure of the ‘golden tide’ seaweeds, Sargassum horneri, in Korean waters
Article Snippet: Polymerase chain reaction (PCR) amplification was carried out in a reaction volume of 15 μl containing 1 × PCR buffer, 25 μM of each dNTP (Bio Basic Inc., Markham, ON, Canada), 0.6 μM of each of the forward and reverse primers, 0.2 units of Taq DNA polymerase (Thermo Fisher Scientific, Waltham, MA, USA) and approximately 5−10 ng of genomic DNA. .. The purified mtDNA fragments were subjected to direct sequencing only in the reverse direction using the same reverse primers as in the PCR and the BigDye Terminator 3.1 Cycle Sequencing Ready Reaction Kit in an ABI 3730xl automated DNA sequencer (Applied Biosystems, Foster City, CA, USA).

Article Title: Exhausting exercise and tissue-specific expression of monocarboxylate transporters in rainbow trout
Article Snippet: Paragraph title: Cloning, sequencing, and partial characterization of MCTs. ... After cDNA synthesis, a PCR reaction was performed as follows: 1× PCR buffer, 1.5 mM MgSO4 , 0.2 mM dNTP, 0.2 mM of Zfish-MCT F and Zfish-MCT R each, 2 μl cDNA, 2 U Taq polymerase (Bio Basic; Amherst, NY), and DNase/RNase-free water for a final volume of 50 μl.

Article Title: Cis-acting requirements in flanking DNA for the programmed elimination of mse2.9: a common mechanism for deletion of internal eliminated sequences from the developing macronucleus of Tetrahymena thermophila
Article Snippet: These primer sets amplify junctions only in transformed strains, as 3r and 5r are complementary to rDNA sequence. .. PCR was performed in a 50 µl vol containing 0.5 µM each primer, 0.2 mM each dNTP, 10× tsg buffer, 1.5 mM MgCl2 and 2.5 U tsg (Biobasic, Toronto, Canada).

Article Title: The role of structural pleiotropy and regulatory evolution in the retention of heteromers of paralogs
Article Snippet: We confirmed the insertion of the DHFR fragments at the correct location by colony PCR using a specific forward Oligo-C targeting a few hundred base pairs upstream of the fusion and a reverse complement oligonucleotide ADHterm_R located in the ADH terminator after the DHFR fragment sequence ( Table S12). .. Tubes were centrifuged for 5 min at 1800 g and 2.5 µL of supernatant was added to a PCR mix composed of 16.85 µL of DNAse free water, 2.5 µL of 10X Taq buffer (BioShop Canada Inc, Canada), 1.5 µL of 25 mM MgCl2, 0.5 µL of 10 mM dNTP (Bio Basic Inc, Canada), 0.15 µL of 5 U/µL Taq DNA polymerase (BioShop Canada Inc, Canada), 0.5 µL of 10 µM Oligo-C and 0.5 µL of 10 µM ADHterm_R.

Article Title: Contrasting life histories contribute to divergent patterns of genetic diversity and population connectivity in freshwater sculpin fishes
Article Snippet: Paragraph title: MtDNA control region sequencing ... Polymerase chain reaction (PCR) amplification was performed in a reaction volume of 15 μl comprising 1× PCR buffer, 25 μM of each dNTP (Bio Basic Inc., Markham, ON, Canada), 0.6 μM of each of the forward and reverse primers and 0.2 U of Taq polymerase (Thermo Fisher Scientific, Waltham, MA, USA).

Construct:

Article Title: Cis-acting requirements in flanking DNA for the programmed elimination of mse2.9: a common mechanism for deletion of internal eliminated sequences from the developing macronucleus of Tetrahymena thermophila
Article Snippet: PCR was used to amplify junction sequences of processing constructs obtained by transformation. .. PCR was performed in a 50 µl vol containing 0.5 µM each primer, 0.2 mM each dNTP, 10× tsg buffer, 1.5 mM MgCl2 and 2.5 U tsg (Biobasic, Toronto, Canada).

Article Title: The role of structural pleiotropy and regulatory evolution in the retention of heteromers of paralogs
Article Snippet: DHFR strains We identified 485 pairs of SSDs and 156 pairs of WGDs present in the Yeast Protein Interactome Collection ( ) and another set of 155 strains constructed by . .. Tubes were centrifuged for 5 min at 1800 g and 2.5 µL of supernatant was added to a PCR mix composed of 16.85 µL of DNAse free water, 2.5 µL of 10X Taq buffer (BioShop Canada Inc, Canada), 1.5 µL of 25 mM MgCl2, 0.5 µL of 10 mM dNTP (Bio Basic Inc, Canada), 0.15 µL of 5 U/µL Taq DNA polymerase (BioShop Canada Inc, Canada), 0.5 µL of 10 µM Oligo-C and 0.5 µL of 10 µM ADHterm_R.

Polyacrylamide Gel Electrophoresis:

Article Title: Multiple Mating and Family Structure of the Western Tent Caterpillar, Malacosoma californicum pluviale: Impact on Disease Resistance
Article Snippet: PCR reactions for all microsatellites used 10 ng of genomic DNA, 0.0625 mM dNTP, (1X) PCR Reaction buffer (Bio Basic Inc. ON, Canada), 0.25 units of Taq (Bio Basics Inc.), 0.75 or 1 mM MgCl2 (Fermentas, ON, Canada; optimized for each locus, see ), 0.15 pmol of fluorescently labeled forward primer (IRDye® 700 or 800, Integrated DNA Technologies, CA, USA), 3 pmol reverse primer (Eurofins MGW Operon, Huntsville, AL, USA), 6.15 or 6.25 μL H2 0 for a total reaction volume of 10 μL. .. Amplified products were denatured and run on a 6% polyacrylamide gel electrophoresis for 1.5 hours on a LI-COR 4300 automated sequencer (LICOR Inc., NE, USA) with a minimum of four size standards (50–350 bp or 50–700 bp LICOR Inc.) per 64 well gel and two to four reference samples.

Mouse Assay:

Article Title: Expression of Nesfatin-1/NUCB2 in Fetal, Neonatal and Adult Mice
Article Snippet: RNA extraction and cDNA synthesis Mice were euthanized by CO2 anesthesia followed by cervical dislocation. .. First strand cDNA synthesis was performed using 2 μg RNA, 10 pmol oligo dT and RNase-free DEPC solution at 70°C for 5 min, followed by double-strand synthesis in 5X RT buffer (Invitrogen, Carlsbad, CA) with 8 mM dNTP (BIO BASIC INC., Ontario, Canada), 200 unit/μl RTase (Invitrogen, Carlsbad, CA) and RNase- free DEPC solution at 37°C for 60min and at 72°C for 15 min.

Plasmid Preparation:

Article Title: Exhausting exercise and tissue-specific expression of monocarboxylate transporters in rainbow trout
Article Snippet: After cDNA synthesis, a PCR reaction was performed as follows: 1× PCR buffer, 1.5 mM MgSO4 , 0.2 mM dNTP, 0.2 mM of Zfish-MCT F and Zfish-MCT R each, 2 μl cDNA, 2 U Taq polymerase (Bio Basic; Amherst, NY), and DNase/RNase-free water for a final volume of 50 μl. .. The amplicons [565 base pair (bp)] obtained from both brain and white muscle were subcloned into TopoTA vector (Invitrogen) and transformed in chemically competent Escherichia coli cells.

Software:

Article Title: Environmental and genetic determinants of transcriptional plasticity in Chinook salmon
Article Snippet: PCR reactions consisted of 20 mM Tris-HCl pH 8.75, 10 mM KCl, 10 mM (NH4 )2 SO4 , 0.1% Triton X-100, 0.1 mg/mL BSA, 200 μM each dNTP, 200 nM forward and reverse primers, 2.0 mM MgSO4 , 0.5 U of taq polymerase (Bio Basic Canada Inc., Markham, ON) and ~50 ng of DNA. .. Conditions for thermal cycling were 95 °C for 2 min, 35 cycles of 95 °C for 15 s, locus-specific annealing temperature (52 °C–OtsG68, 56 °C–OtsG432, 58 °C–Ots208, 60 °C–Ots209, Ots211) for 15 s and 72 °C for 30 s, followed by 72 °C for 5 min. Microsatellite PCR products were characterized using a Licor 4300 DNA Analyzer (Licor Biosciences Inc.) and Gene ImagR software (Scanalytics Inc.).

SYBR Green Assay:

Article Title: Tissue-Specific Localization NUCB2/nesfatin-1 in the Liver and Heart of Mouse Fetus
Article Snippet: .. First strand cDNA synthesis was performed using 2 μg RNA, 10 pmol oligo dT and RNase-free DEPC solution at 70° for 5min, followed by double-strand synthesis in 5× RT buffer (Invitrogen, Carlsbad, CA, USA) with 8 mM dNTP (BIO BASIC INC., Ontario, Canada), 200 unit/μL RTase (Invitrogen, Carlsbad, CA, USA) and RNase-free DEPC solution at 37° for 60 min and at 72° for 15 min. qRT-PCR was performed in a total volume of 20 μL buffer solution containing 2 μL of template cDNA, 10 μL of SYBR Green (Roche, Manheim, Germany), and 10 pmol of each primer. ..

Article Title: Calorie restriction effects on circadian rhythms in gene expression are sex dependent
Article Snippet: The RT mix was prepared using 1 µg of mRNA, 50 ng of random hexamer (N808127, Invitrogen), 10 mM dNTP (DD0058, Biobasic), 0.1 M DDT and RNaseOUT Recombinant RNase inhibitor (10777-019, 40 units/ µl) and it was reverse transcribed using SuperScript III Reverse Transcriptase (18080-044, Invitrogen). .. RNA quantification was performed using qPCR with Universal SYBR Green Mix (1725125, BioRad).

RNA Extraction:

Article Title: Expression of Nesfatin-1/NUCB2 in Fetal, Neonatal and Adult Mice
Article Snippet: Paragraph title: RNA extraction and cDNA synthesis ... First strand cDNA synthesis was performed using 2 μg RNA, 10 pmol oligo dT and RNase-free DEPC solution at 70°C for 5 min, followed by double-strand synthesis in 5X RT buffer (Invitrogen, Carlsbad, CA) with 8 mM dNTP (BIO BASIC INC., Ontario, Canada), 200 unit/μl RTase (Invitrogen, Carlsbad, CA) and RNase- free DEPC solution at 37°C for 60min and at 72°C for 15 min.

Article Title: Th 17 Cells and Nesfatin-1 are associated with Spontaneous Abortion in the CBA/j × DBA/2 Mouse Model
Article Snippet: Paragraph title: RNA extraction and cDNA synthesis ... First strand cDNA synthesis was performed in RNase-free DEPC solution containing 2 μg total RNA and 10 pmol oligo dT at 70°C for 5 min, followed by double-strand synthesis in 5× RT buffer (Invitrogen, Carlsbad, CA) with 8 mM dNTP (BIO BASIC INC., Ontario, Canada), 200 unit/μL RTase (Invitrogen, Carlsbad, CA) at 37°C for 60min and at 72°C for 15 min.

Article Title: Tissue-Specific Localization NUCB2/nesfatin-1 in the Liver and Heart of Mouse Fetus
Article Snippet: Paragraph title: RNA extraction and qRT-PCR ... First strand cDNA synthesis was performed using 2 μg RNA, 10 pmol oligo dT and RNase-free DEPC solution at 70° for 5min, followed by double-strand synthesis in 5× RT buffer (Invitrogen, Carlsbad, CA, USA) with 8 mM dNTP (BIO BASIC INC., Ontario, Canada), 200 unit/μL RTase (Invitrogen, Carlsbad, CA, USA) and RNase-free DEPC solution at 37° for 60 min and at 72° for 15 min. qRT-PCR was performed in a total volume of 20 μL buffer solution containing 2 μL of template cDNA, 10 μL of SYBR Green (Roche, Manheim, Germany), and 10 pmol of each primer.

Binding Assay:

Article Title: Environmental and genetic determinants of transcriptional plasticity in Chinook salmon
Article Snippet: DNA was extracted from a small piece of fin tissue (parents and offspring) using a silica binding-column procedure (Elphinstone et al. ). .. PCR reactions consisted of 20 mM Tris-HCl pH 8.75, 10 mM KCl, 10 mM (NH4 )2 SO4 , 0.1% Triton X-100, 0.1 mg/mL BSA, 200 μM each dNTP, 200 nM forward and reverse primers, 2.0 mM MgSO4 , 0.5 U of taq polymerase (Bio Basic Canada Inc., Markham, ON) and ~50 ng of DNA.

Agarose Gel Electrophoresis:

Article Title: Exhausting exercise and tissue-specific expression of monocarboxylate transporters in rainbow trout
Article Snippet: The RNA was quantified using NanoDrop 2000 (Thermo Scientific; Wilmington, DE), and the purity and integrity were verified on a 1.5% agarose gel prepared with 1× MOPS buffer. cDNA synthesis was performed using 5 μg of total RNA, 200 U SuperScript II reverse transcriptase, 250 ng random octamers (Invitrogen), and DNase/RNase-free water for a total volume of 20 μl. .. After cDNA synthesis, a PCR reaction was performed as follows: 1× PCR buffer, 1.5 mM MgSO4 , 0.2 mM dNTP, 0.2 mM of Zfish-MCT F and Zfish-MCT R each, 2 μl cDNA, 2 U Taq polymerase (Bio Basic; Amherst, NY), and DNase/RNase-free water for a final volume of 50 μl.

Article Title: Population genetic structure of eelgrass (Zostera marina) on the Korean coast: Current status and conservation implications for future management
Article Snippet: PCR amplification was accomplished in a reaction volume of 15 μl containing 25 μM of each dNTP (Bio Basic), 0.6 μM each of the forward and reverse primers, 0.2 units of Taq DNA polymerase (Thermo Fisher Scientific), 1× PCR buffer, and approximately 5−10 ng of template DNA. .. Each PCR product was checked on a 2% agarose gel stained with RedsafeTM (iNtRON Biotechnology).

Article Title: Calorie restriction effects on circadian rhythms in gene expression are sex dependent
Article Snippet: The RNA pellet was dissolved in water and quantified on Nanodrop, RNA quality was checked on 1% agarose gel. .. The RT mix was prepared using 1 µg of mRNA, 50 ng of random hexamer (N808127, Invitrogen), 10 mM dNTP (DD0058, Biobasic), 0.1 M DDT and RNaseOUT Recombinant RNase inhibitor (10777-019, 40 units/ µl) and it was reverse transcribed using SuperScript III Reverse Transcriptase (18080-044, Invitrogen).

Quantitation Assay:

Article Title: Functional Evaluation of Genetic and Environmental Regulators of P450 mRNA Levels
Article Snippet: Paragraph title: Absolute Quantitation of mRNA level by Real-time PCR ... Reverse transcription reactions were prepared in a total volume of 20 µl containing 1× RT buffer, 0.5 mM dNTP, 0.5 µM poly24dT, 5 µM N9, 20 U RNase inhibitor (BIO BASIC INC.) and 100 U M-MLV(H-) reverse transcriptase (Promega).

Concentration Assay:

Article Title: Population genetic structure of eelgrass (Zostera marina) on the Korean coast: Current status and conservation implications for future management
Article Snippet: DNA concentration was determined using a Qubit® 2.0 Fluorometer (Invitrogen). .. PCR amplification was accomplished in a reaction volume of 15 μl containing 25 μM of each dNTP (Bio Basic), 0.6 μM each of the forward and reverse primers, 0.2 units of Taq DNA polymerase (Thermo Fisher Scientific), 1× PCR buffer, and approximately 5−10 ng of template DNA.

Marker:

Article Title: The role of structural pleiotropy and regulatory evolution in the retention of heteromers of paralogs
Article Snippet: Tubes were centrifuged for 5 min at 1800 g and 2.5 µL of supernatant was added to a PCR mix composed of 16.85 µL of DNAse free water, 2.5 µL of 10X Taq buffer (BioShop Canada Inc, Canada), 1.5 µL of 25 mM MgCl2, 0.5 µL of 10 mM dNTP (Bio Basic Inc, Canada), 0.15 µL of 5 U/µL Taq DNA polymerase (BioShop Canada Inc, Canada), 0.5 µL of 10 µM Oligo-C and 0.5 µL of 10 µM ADHterm_R. .. The DHFR fragments and associated resistance modules were amplified from plasmids pAG25-linker-F[1,2]-ADHterm (NAT resistance marker) and pAG32-linker-F[3]-ADHterm (HygB resistance marker) ( ) using oligonucleotides defined in ( Table S12).

Staining:

Article Title: The origin and population genetic structure of the ‘golden tide’ seaweeds, Sargassum horneri, in Korean waters
Article Snippet: Polymerase chain reaction (PCR) amplification was carried out in a reaction volume of 15 μl containing 1 × PCR buffer, 25 μM of each dNTP (Bio Basic Inc., Markham, ON, Canada), 0.6 μM of each of the forward and reverse primers, 0.2 units of Taq DNA polymerase (Thermo Fisher Scientific, Waltham, MA, USA) and approximately 5−10 ng of genomic DNA. .. RCR thermal cycling conditions comprised an initial denaturation at 94 °C for 3 min, followed by 35 cycles of denaturation at 94 °C for 20 s, annealing at 50 °C for 50 s and extension at 68 °C for 60 s, and a final extension at 68 °C for 10 min. PCR products were checked on 2% agarose gels stained with Redsafe (iNtRON).

Article Title: Population genetic structure of eelgrass (Zostera marina) on the Korean coast: Current status and conservation implications for future management
Article Snippet: PCR amplification was accomplished in a reaction volume of 15 μl containing 25 μM of each dNTP (Bio Basic), 0.6 μM each of the forward and reverse primers, 0.2 units of Taq DNA polymerase (Thermo Fisher Scientific), 1× PCR buffer, and approximately 5−10 ng of template DNA. .. Each PCR product was checked on a 2% agarose gel stained with RedsafeTM (iNtRON Biotechnology).

Article Title: Contrasting life histories contribute to divergent patterns of genetic diversity and population connectivity in freshwater sculpin fishes
Article Snippet: Polymerase chain reaction (PCR) amplification was performed in a reaction volume of 15 μl comprising 1× PCR buffer, 25 μM of each dNTP (Bio Basic Inc., Markham, ON, Canada), 0.6 μM of each of the forward and reverse primers and 0.2 U of Taq polymerase (Thermo Fisher Scientific, Waltham, MA, USA). .. PCR products were checked on 2% agarose gels stained with RedSafe (iNtRon Biotechnology, Daejeon, Korea).

Fluorescence In Situ Hybridization:

Article Title: Exhausting exercise and tissue-specific expression of monocarboxylate transporters in rainbow trout
Article Snippet: Therefore, total RNA (from two fish not used in the gene expression experiments) was isolated from white muscle and brain using the TRIzol method (Invitrogen; Carlsbad, CA). .. After cDNA synthesis, a PCR reaction was performed as follows: 1× PCR buffer, 1.5 mM MgSO4 , 0.2 mM dNTP, 0.2 mM of Zfish-MCT F and Zfish-MCT R each, 2 μl cDNA, 2 U Taq polymerase (Bio Basic; Amherst, NY), and DNase/RNase-free water for a final volume of 50 μl.

Article Title: Environmental and genetic determinants of transcriptional plasticity in Chinook salmon
Article Snippet: Because we combined multiple families in each semi-natural enclosure, we used microsatellite genotypes to assign parentage for all the fish sampled from the semi-natural enclosures. .. PCR reactions consisted of 20 mM Tris-HCl pH 8.75, 10 mM KCl, 10 mM (NH4 )2 SO4 , 0.1% Triton X-100, 0.1 mg/mL BSA, 200 μM each dNTP, 200 nM forward and reverse primers, 2.0 mM MgSO4 , 0.5 U of taq polymerase (Bio Basic Canada Inc., Markham, ON) and ~50 ng of DNA.

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