Total RNA of cells (106) was extracted with Trizol and was reverse transcribed with PrimeScriptTM RT Maseter Mix (Takara, RR036A). Real-time PCR was performed with SYBR Premix Ex TaqTM IIz (Takara, RR820A), using a ROCHE LightCycler® 480. The samples were analyzed using the 2-△△Ct method from the Ct values of the respective RNAs (PKG, cGMP, and CYP11B2) relative to the housekeeping gene GAPDH. The following primers were used. cGMP primer (sense: 5′-ATTGGGATTGTGGGCCATGT-3′; antisense: 5′-TTGTCACGTAGTCGGTGAGC-3′); PKG primer (sense: 5′-CGTTACCCGAGAAGACTCACC-3′; antisense: 5′-GAGACATCATCCAGTCCTCCAA-3′); CYP11B2 primer (sense: 5′-TGCTAAATGGGGCTGACTGG-3′; antisense: 5′-CATCCATGGTAAGGCTCCCTC-3′).
Primescript rt maseter mix
Manufactured by Takara Bio
PrimeScript RT Master Mix is a reverse transcription reagent kit used for the synthesis of first-strand cDNA from RNA templates. It contains high-quality reverse transcriptase, RNase inhibitor, and other necessary components for efficient cDNA synthesis.
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Lab products found in correlation
3 protocols using primescript rt maseter mix
1
Real-time PCR Analysis of Signaling Pathways
2
Real-time PCR Analysis of sGC, cGMP, and PKG
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Total RNA of cells (106) was extracted with Trizol and was reverse transcribed with PrimeScriptTM RT Maseter Mix (Takara, RR036A). Real-time PCR was performed with SYBR Premix Ex TaqTM IIz (Takara, RR820A), using a ROCHE LightCycler® 480. The samples were analyzed using the 2−△△Ct method from the Ct values of the respective RNAs (sGC, cGMP, and PKG) relative to the housekeeping gene GAPDH. The following primers were used. sGC primer (sense: 5′- ACGGAGCCAATAGAGGAGGT-3′; antisense: 5′- TGCCAGGGTGTGTAGGTAGA-3′); cGMP primer (sense: 5′-ATTGGGATTGTGGGCCATGT-3′; antisense: 5′-TTGTCACGTAGTCGGTGAGC-3′); PKG primer (sense: 5′-CGTTACCCGAGAAGACTCACC-3′; antisense: 5′-GAGACATCATCCAGTCCTCCAA-3′).
3
Analyzing Progerin and Oxidative Stress Markers in Blood Granulocytes
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Total RNA of the peripheral blood granulocytes of patients and rat primary LSECs (106) was extracted with Trizol and reverse-transcribed with PrimeScriptTM RT Maseter Mix (Takara, #RR036A). Real-time PCR was performed with SYBR Premix Ex TaqTM IIz (Takara, #RR820A), using ROCHE LightCycler®480. The samples were analyzed using the 2-∆∆Ct method from the Ct values of the respective RNAs (progerin, NOX2, NOX4, LMNA, Lamin B1, and Cav-1) relative to the housekeeping gene GAPDH. The primer sequences are displayed in the Supplementary Materials . The experiment was repeated more than three times.
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