Fluorescence:Article Title: RNAi-induced knockdown of white gene in the southern green stink bug ( Nezara viridula L.)
Article Snippet: .. Similarly, a non-specific green fluorescence protein (GFP) gene fragment of 503 bp was amplified from pGLOTM plasmid (Bio-Rad, Hercules, CA, Cat. No. 1660405EDU) by PCR using the forward (5′-T7- AGGTGATGCTACATACGGAAAG-3′) and reverse (5′-T7-ACAGGTAATGGTTGTCTGGTAAA-3′) primers. .. The PCR reactions were carried out using a C1000 TouchTM Thermal Cycler (Bio-Rad Laboratories, Singapore) and the temperature profiles included an initial heating step at 94 °C for 2 min, followed by 39 cycles of denaturation at 94 °C for 15 s, annealing at 60 °C for 30 s, and extension at 68 °C for 30 s. PCR products generated were then used for dsRNA synthesis using MEGAscriptTM RNAi Kit (InvitrogenTM by Thermo Fischer ScientificTM, Vilnius, LT, Cat. No. AM1626) according to manufacturer instructions ( https://assets.thermofisher.com/TFS-Assets/LSG/manuals/cms_072987.pdf ).
Amplification:Article Title: RNAi-induced knockdown of white gene in the southern green stink bug ( Nezara viridula L.)
Article Snippet: .. Similarly, a non-specific green fluorescence protein (GFP) gene fragment of 503 bp was amplified from pGLOTM plasmid (Bio-Rad, Hercules, CA, Cat. No. 1660405EDU) by PCR using the forward (5′-T7- AGGTGATGCTACATACGGAAAG-3′) and reverse (5′-T7-ACAGGTAATGGTTGTCTGGTAAA-3′) primers. .. The PCR reactions were carried out using a C1000 TouchTM Thermal Cycler (Bio-Rad Laboratories, Singapore) and the temperature profiles included an initial heating step at 94 °C for 2 min, followed by 39 cycles of denaturation at 94 °C for 15 s, annealing at 60 °C for 30 s, and extension at 68 °C for 30 s. PCR products generated were then used for dsRNA synthesis using MEGAscriptTM RNAi Kit (InvitrogenTM by Thermo Fischer ScientificTM, Vilnius, LT, Cat. No. AM1626) according to manufacturer instructions ( https://assets.thermofisher.com/TFS-Assets/LSG/manuals/cms_072987.pdf ).
Plasmid Preparation:Article Title: RNAi-induced knockdown of white gene in the southern green stink bug ( Nezara viridula L.)
Article Snippet: .. Similarly, a non-specific green fluorescence protein (GFP) gene fragment of 503 bp was amplified from pGLOTM plasmid (Bio-Rad, Hercules, CA, Cat. No. 1660405EDU) by PCR using the forward (5′-T7- AGGTGATGCTACATACGGAAAG-3′) and reverse (5′-T7-ACAGGTAATGGTTGTCTGGTAAA-3′) primers. .. The PCR reactions were carried out using a C1000 TouchTM Thermal Cycler (Bio-Rad Laboratories, Singapore) and the temperature profiles included an initial heating step at 94 °C for 2 min, followed by 39 cycles of denaturation at 94 °C for 15 s, annealing at 60 °C for 30 s, and extension at 68 °C for 30 s. PCR products generated were then used for dsRNA synthesis using MEGAscriptTM RNAi Kit (InvitrogenTM by Thermo Fischer ScientificTM, Vilnius, LT, Cat. No. AM1626) according to manufacturer instructions ( https://assets.thermofisher.com/TFS-Assets/LSG/manuals/cms_072987.pdf ).
Polymerase Chain Reaction:Article Title: RNAi-induced knockdown of white gene in the southern green stink bug ( Nezara viridula L.)
Article Snippet: .. Similarly, a non-specific green fluorescence protein (GFP) gene fragment of 503 bp was amplified from pGLOTM plasmid (Bio-Rad, Hercules, CA, Cat. No. 1660405EDU) by PCR using the forward (5′-T7- AGGTGATGCTACATACGGAAAG-3′) and reverse (5′-T7-ACAGGTAATGGTTGTCTGGTAAA-3′) primers. .. The PCR reactions were carried out using a C1000 TouchTM Thermal Cycler (Bio-Rad Laboratories, Singapore) and the temperature profiles included an initial heating step at 94 °C for 2 min, followed by 39 cycles of denaturation at 94 °C for 15 s, annealing at 60 °C for 30 s, and extension at 68 °C for 30 s. PCR products generated were then used for dsRNA synthesis using MEGAscriptTM RNAi Kit (InvitrogenTM by Thermo Fischer ScientificTM, Vilnius, LT, Cat. No. AM1626) according to manufacturer instructions ( https://assets.thermofisher.com/TFS-Assets/LSG/manuals/cms_072987.pdf ).
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