Total RNA from HL-1 cells and atrial tissues was acquired using TRIzol reagent following the manufacturer’s instructions (TIANGEN, DP421). Complementary DNA was synthesized with TIANScript complementary DNA kits (TIANGEN, KR104) and real-time PCR with a Cycler Sequence Detection System (Bio-Rad, CFX96). The PCR cycle was as follows: 40 cycles at 95 °C for 10 s, 55 °C for 20 s, and 72 °C for 20 s. The relative mRNA levels were assessed using the 2−ΔΔCt method and normalized to actb. The primer sequences are available in Table S2 .
Cycler sequence detection system
Manufactured by Bio-Rad
Sourced in United States
The Cycler Sequence Detection System is a laboratory instrument designed for real-time PCR (polymerase chain reaction) analysis. It combines thermal cycling capabilities with fluorescence detection to enable the amplification and quantification of DNA or RNA sequences.
Automatically generated - may contain errors
2 protocols using cycler sequence detection system
1
Real-time PCR analysis of mRNA levels
2
Quantitative Analysis of Gene Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Total RNA was extracted from macrophages or tissues using the TRIzol reagent (Sigma-Aldrich, T9424). Complementary DNA (cDNA) was synthesized using the TIANScript cDNA kits (TIANGEN, KR104). Then, cDNA was amplified using a SYBR Green PCR kit (TIANGEN, FP402). qRT-PCR analysis was performed using a Cycler Sequence Detection System (Bio-Rad, CFX96, USA). The cycling parameters were as follows: 40 cycles at 95 °C for 15 s, 55 °C for 15 s and 72 °C for 15 s. Relative expression of target genes normalized to Gapdh were analyzed with CFX Manager 2.1 software (Bio-Rad).
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