Amplification:Article Title: Non-Redundant and Overlapping Oncogenic Readouts of Non-Canonical and Novel Colorectal Cancer KRAS and NRAS Mutants
Article Snippet: .. The mutant KRAS and NRAS variant fragments were amplified in a polymerase chain reaction (PCR) mixture containing a final concentration of 1X PCR buffer (Titanium® Taq PCR buffer, Clontech Laboratories, Inc., Mountainview, CA, USA), 0.125 μM of each deoxynucleoside triphosphate (Promega, Madison, WI, USA), 2 μM each of the forward and reverse primers, 1U Taq polymerase (Titanium® Taq polymerase, Clontech Laboratories, Inc.), and 50 ng of the pTargeT-WT KRAS or pTargeT-WT NRAS template. .. Amplification was performed with an initial denaturation temperature of 94 °C for 5 min, followed by 25–30 cycles of denaturation at 94 °C for 30 s, annealing at 55 °C for 30 s, and extension at 72 °C for 30 s, with a final extension step at 72 °C for 10 min.
Mutagenesis:Article Title: Non-Redundant and Overlapping Oncogenic Readouts of Non-Canonical and Novel Colorectal Cancer KRAS and NRAS Mutants
Article Snippet: .. The mutant KRAS and NRAS variant fragments were amplified in a polymerase chain reaction (PCR) mixture containing a final concentration of 1X PCR buffer (Titanium® Taq PCR buffer, Clontech Laboratories, Inc., Mountainview, CA, USA), 0.125 μM of each deoxynucleoside triphosphate (Promega, Madison, WI, USA), 2 μM each of the forward and reverse primers, 1U Taq polymerase (Titanium® Taq polymerase, Clontech Laboratories, Inc.), and 50 ng of the pTargeT-WT KRAS or pTargeT-WT NRAS template. .. Amplification was performed with an initial denaturation temperature of 94 °C for 5 min, followed by 25–30 cycles of denaturation at 94 °C for 30 s, annealing at 55 °C for 30 s, and extension at 72 °C for 30 s, with a final extension step at 72 °C for 10 min.
Purification:Article Title: Chemical, Microbiological, and Functional Characterization of Kefir Produced from Cow's Milk and Soy Milk
Article Snippet: .. A total of 30 μ L polymerase chain reaction (PCR) mixture contained 3 μ L buffer solution, 2.4 μ L dNTPs, 1.5 μ L of each primer, 0.15 μ L Ex Taq polymerase (Takara, Shiga, Japan), 20.85 μ L purified water, and 0.6 μ L of the extracted DNA. .. The PCR amplification was carried out as follows: for bacteria, template DNA was denatured for 2 min at 96°C, followed by 25 cycles of denaturing at 96°C for 15 s, annealing at 50°C for 15 s, and primer extension at 72°C for 1 min 30 s; for yeasts, template DNA was denatured for 2 min at 96°C, followed by 25 cycles of denaturing at 96°C for 15 s, annealing at 50°C for 15 s, and primer extension at 72°C for 40 s. The amplification products were analysed by electrophoresis on 1.0% agarose gels before being sequenced by Eurofins Genomics Co. Ltd. (Tokyo, Japan).
SYBR Green Assay:Article Title: Applicability of Hyaluronic Acid-Alginate Hydrogel and Ovarian Cells for In Vitro Development of Mouse Preantral Follicles
Article Snippet: .. The polymerase chain reaction (PCR) mix for each well contained 5 ml Power SYBR Green PCR Master Mix (Takara, Japan), 1 ml dH2 O, 1 ml of each of the forward and reverse primers (5 pmol/ml), and 2 ml of single-strand cDNA in a final reaction volume of 10 ml. .. PCR was performed on the ABI StepOnePlus real-time PCR system (Applied Biosystems) using the following program: stage 1: 95˚C for 10 minutes; stage 2: 40 cycles of 95˚C for 15 seconds and 60˚C for 1 minute; and stage 3: 95˚C for 15 seconds, 60˚C for 15 seconds, and 95˚C for 15 seconds.
Concentration Assay:Article Title: Assessment and molecular characterization of Bacillus cereus isolated from edible fungi in China
Article Snippet: .. The polymerase chain reaction (PCR) mixture (25 μL) comprised of 50 ng genomic DNA, 0.5 μL of each primer solution (concentration 10 μM), and 12.5 μL PCR Premix Taq™ (Takara, China). .. The amplification process included: initial denaturation at 94 °C for 5 min, 30 cycles of 94 °C for 1 min, 55 °C or 58 °C (58 °C for cesB and 55 °C for the other toxin genes) for 1 min, 72 °C for 1 min, and a final 10 min extension at 72 °C [ , , , ].
Article Title: Non-Redundant and Overlapping Oncogenic Readouts of Non-Canonical and Novel Colorectal Cancer KRAS and NRAS Mutants
Article Snippet: .. The mutant KRAS and NRAS variant fragments were amplified in a polymerase chain reaction (PCR) mixture containing a final concentration of 1X PCR buffer (Titanium® Taq PCR buffer, Clontech Laboratories, Inc., Mountainview, CA, USA), 0.125 μM of each deoxynucleoside triphosphate (Promega, Madison, WI, USA), 2 μM each of the forward and reverse primers, 1U Taq polymerase (Titanium® Taq polymerase, Clontech Laboratories, Inc.), and 50 ng of the pTargeT-WT KRAS or pTargeT-WT NRAS template. .. Amplification was performed with an initial denaturation temperature of 94 °C for 5 min, followed by 25–30 cycles of denaturation at 94 °C for 30 s, annealing at 55 °C for 30 s, and extension at 72 °C for 30 s, with a final extension step at 72 °C for 10 min.
Polymerase Chain Reaction:Article Title: Genetic variability of the Aedes aegypti (Diptera: Culicidae) mosquito in El Salvador, vector of dengue, yellow fever, chikungunya and Zika
Article Snippet: .. A polymerase chain reaction (PCR) mix for six samples consisted of the following: 195.6 μl sterile ultra-pure water; 2.4 μl Taq polymerase (Clonetech, Mountainview, CA, USA); 30 μl Taq 10× buffer; 24 μl dNTPs; 6 μl forward primer; and 6 μl reverse primer and 2 μl template DNA (~100 ng template). ..
Article Title: Chemical, Microbiological, and Functional Characterization of Kefir Produced from Cow's Milk and Soy Milk
Article Snippet: .. A total of 30 μ L polymerase chain reaction (PCR) mixture contained 3 μ L buffer solution, 2.4 μ L dNTPs, 1.5 μ L of each primer, 0.15 μ L Ex Taq polymerase (Takara, Shiga, Japan), 20.85 μ L purified water, and 0.6 μ L of the extracted DNA. .. The PCR amplification was carried out as follows: for bacteria, template DNA was denatured for 2 min at 96°C, followed by 25 cycles of denaturing at 96°C for 15 s, annealing at 50°C for 15 s, and primer extension at 72°C for 1 min 30 s; for yeasts, template DNA was denatured for 2 min at 96°C, followed by 25 cycles of denaturing at 96°C for 15 s, annealing at 50°C for 15 s, and primer extension at 72°C for 40 s. The amplification products were analysed by electrophoresis on 1.0% agarose gels before being sequenced by Eurofins Genomics Co. Ltd. (Tokyo, Japan).
Article Title: Applicability of Hyaluronic Acid-Alginate Hydrogel and Ovarian Cells for In Vitro Development of Mouse Preantral Follicles
Article Snippet: .. The polymerase chain reaction (PCR) mix for each well contained 5 ml Power SYBR Green PCR Master Mix (Takara, Japan), 1 ml dH2 O, 1 ml of each of the forward and reverse primers (5 pmol/ml), and 2 ml of single-strand cDNA in a final reaction volume of 10 ml. .. PCR was performed on the ABI StepOnePlus real-time PCR system (Applied Biosystems) using the following program: stage 1: 95˚C for 10 minutes; stage 2: 40 cycles of 95˚C for 15 seconds and 60˚C for 1 minute; and stage 3: 95˚C for 15 seconds, 60˚C for 15 seconds, and 95˚C for 15 seconds.
Article Title: Assessment and molecular characterization of Bacillus cereus isolated from edible fungi in China
Article Snippet: .. The polymerase chain reaction (PCR) mixture (25 μL) comprised of 50 ng genomic DNA, 0.5 μL of each primer solution (concentration 10 μM), and 12.5 μL PCR Premix Taq™ (Takara, China). .. The amplification process included: initial denaturation at 94 °C for 5 min, 30 cycles of 94 °C for 1 min, 55 °C or 58 °C (58 °C for cesB and 55 °C for the other toxin genes) for 1 min, 72 °C for 1 min, and a final 10 min extension at 72 °C [ , , , ].
Article Title: Non-Redundant and Overlapping Oncogenic Readouts of Non-Canonical and Novel Colorectal Cancer KRAS and NRAS Mutants
Article Snippet: .. The mutant KRAS and NRAS variant fragments were amplified in a polymerase chain reaction (PCR) mixture containing a final concentration of 1X PCR buffer (Titanium® Taq PCR buffer, Clontech Laboratories, Inc., Mountainview, CA, USA), 0.125 μM of each deoxynucleoside triphosphate (Promega, Madison, WI, USA), 2 μM each of the forward and reverse primers, 1U Taq polymerase (Titanium® Taq polymerase, Clontech Laboratories, Inc.), and 50 ng of the pTargeT-WT KRAS or pTargeT-WT NRAS template. .. Amplification was performed with an initial denaturation temperature of 94 °C for 5 min, followed by 25–30 cycles of denaturation at 94 °C for 30 s, annealing at 55 °C for 30 s, and extension at 72 °C for 30 s, with a final extension step at 72 °C for 10 min.
Article Title: A Comprehensive Census of Microbial Diversity in Hot Springs of Tengchong, Yunnan Province China Using 16S rRNA Gene Pyrosequencing
Article Snippet: .. The polymerase chain reaction (PCR) mix contained 10 ng of template DNA, 5 µl rTaq reaction buffer, 400 nM of each primer, 200 µM dNTPs, and 0.3 unit of rTaq polymerase (Takara, Dalian, China) in a 25 µl reaction. .. The amplification procedure was as follows: an initial denaturation step at 95°C for 5 min, and 30 cycles of denaturing at 94°C for 30 s, annealing at 54°C for 30 s, and extension at 72°C for 1 min, followed by a final extension step at 72°C for 10 min. Amplicons from four PCR were pooled for each sample.
Article Title: TissueGene-C promotes an anti-inflammatory micro-environment in a rat monoiodoacetate model of osteoarthritis via polarization of M2 macrophages leading to pain relief and structural improvement
Article Snippet: .. The polymerase chain reaction (PCR) mixture was prepared to a final volume of 50 µL, as follows: 1 µL cDNA, 0.2 µM of each primer, 10 µL SYBR Premix Ex Taq (TAKARA Bio, Shiga, Japan). .. Expression levels of the target genes were quantified with the ABI 7900 real-time PCR system (RT-PCR, Applied Biosystems, CA, USA).
Article Title: Effects of Dietary Supplementation With Enterococcus faecium and Clostridium butyricum, Either Alone or in Combination, on Growth and Fecal Microbiota Composition of Post-weaning Pigs at a Commercial Farm
Article Snippet: .. Each 50 μL of polymerase chain reaction (PCR) mixture contained 1 μL of sample DNA, 21 μL of MilliQ® water, 25 μL of 2X MightyAmp® Buffer, 1 μL of forward primer (5 μM), 1 μL of reverse primer (5 μM), and 1 μL (1.25 U) of MightyAmp DNA Polymerase (Takara, Tokyo, Japan). ..
Variant Assay:Article Title: Non-Redundant and Overlapping Oncogenic Readouts of Non-Canonical and Novel Colorectal Cancer KRAS and NRAS Mutants
Article Snippet: .. The mutant KRAS and NRAS variant fragments were amplified in a polymerase chain reaction (PCR) mixture containing a final concentration of 1X PCR buffer (Titanium® Taq PCR buffer, Clontech Laboratories, Inc., Mountainview, CA, USA), 0.125 μM of each deoxynucleoside triphosphate (Promega, Madison, WI, USA), 2 μM each of the forward and reverse primers, 1U Taq polymerase (Titanium® Taq polymerase, Clontech Laboratories, Inc.), and 50 ng of the pTargeT-WT KRAS or pTargeT-WT NRAS template. .. Amplification was performed with an initial denaturation temperature of 94 °C for 5 min, followed by 25–30 cycles of denaturation at 94 °C for 30 s, annealing at 55 °C for 30 s, and extension at 72 °C for 30 s, with a final extension step at 72 °C for 10 min.
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