Concentration Assay:Article Title: SETD5 haploinsufficiency affects mitochondrial compartment in neural cells
Article Snippet: .. Then after chilling the first mix on ice for 5′, a second retronscription mix (4 μl of ImProm II 5 × buffer, 3.8 μl MgCl2 (final concentration 2.5 mM), 1 μl dNTP Mix (final concentration 0.5 mM each dNTP), 1 μl ImProm II reverse transcriptase, 5.2 of nuclease free water) is added to first mix, and then reverse transcription is performed with the following reaction profile: 4° × 30′′ 25° × 5′ 42 × 1 h 70° × 15′ 16 × ∞ After c-DNA synthesis real-time quantitative PCR was performed (RT-qPCR) using CFX 96 Real-Time System (Biorad) thermocycler. .. The real-time TITAN (Bioatlas) mix was composed as follows for each reaction samples: 2 μl of c-DNA 9.8 μl of H2O 3.2 μl of TITAN HotTaq EvaGreen qPCR Mix (Titan hotTaq DNA polymerase, 5 × qPCR buffer E, 12.5 mM MgCl2) The amplification profile used was the following: 95° × 15′ (95° × 15′′, 60° × 20′′, 72° × 20′′) × 40 65° RT-qPCR data were analyzed using CFX Manager software (Biorad) and the differential gene expression was calculated using the ∆∆ct methods.
Reverse Transcription:Article Title: SETD5 haploinsufficiency affects mitochondrial compartment in neural cells
Article Snippet: .. Then after chilling the first mix on ice for 5′, a second retronscription mix (4 μl of ImProm II 5 × buffer, 3.8 μl MgCl2 (final concentration 2.5 mM), 1 μl dNTP Mix (final concentration 0.5 mM each dNTP), 1 μl ImProm II reverse transcriptase, 5.2 of nuclease free water) is added to first mix, and then reverse transcription is performed with the following reaction profile: 4° × 30′′ 25° × 5′ 42 × 1 h 70° × 15′ 16 × ∞ After c-DNA synthesis real-time quantitative PCR was performed (RT-qPCR) using CFX 96 Real-Time System (Biorad) thermocycler. .. The real-time TITAN (Bioatlas) mix was composed as follows for each reaction samples: 2 μl of c-DNA 9.8 μl of H2O 3.2 μl of TITAN HotTaq EvaGreen qPCR Mix (Titan hotTaq DNA polymerase, 5 × qPCR buffer E, 12.5 mM MgCl2) The amplification profile used was the following: 95° × 15′ (95° × 15′′, 60° × 20′′, 72° × 20′′) × 40 65° RT-qPCR data were analyzed using CFX Manager software (Biorad) and the differential gene expression was calculated using the ∆∆ct methods.
Real-time Polymerase Chain Reaction:Article Title: SETD5 haploinsufficiency affects mitochondrial compartment in neural cells
Article Snippet: .. Then after chilling the first mix on ice for 5′, a second retronscription mix (4 μl of ImProm II 5 × buffer, 3.8 μl MgCl2 (final concentration 2.5 mM), 1 μl dNTP Mix (final concentration 0.5 mM each dNTP), 1 μl ImProm II reverse transcriptase, 5.2 of nuclease free water) is added to first mix, and then reverse transcription is performed with the following reaction profile: 4° × 30′′ 25° × 5′ 42 × 1 h 70° × 15′ 16 × ∞ After c-DNA synthesis real-time quantitative PCR was performed (RT-qPCR) using CFX 96 Real-Time System (Biorad) thermocycler. .. The real-time TITAN (Bioatlas) mix was composed as follows for each reaction samples: 2 μl of c-DNA 9.8 μl of H2O 3.2 μl of TITAN HotTaq EvaGreen qPCR Mix (Titan hotTaq DNA polymerase, 5 × qPCR buffer E, 12.5 mM MgCl2) The amplification profile used was the following: 95° × 15′ (95° × 15′′, 60° × 20′′, 72° × 20′′) × 40 65° RT-qPCR data were analyzed using CFX Manager software (Biorad) and the differential gene expression was calculated using the ∆∆ct methods.
|
Bio-Rad is a verified supplier 
Bio-Rad manufactures this product 