Total RNA in renal tissues and SV40-MES-13 cells was extracted with the RNA Simple Total RNA Kit (Tiangen, Beijing, China). cDNA was synthesized with the TIANScript IIRT Kit (Tiangen, Beijing, China) at 42 °C for 60 min. qRT-PCR was performed in triplicate with FastFire qPCR PreMix (SYBR Green) (Tiangen, Beijing, China) on a ViPlex Fluor real-time PCR system (Vivantis, Selangor, Malaysia). Thermocycling conditions were set as follows: 95 °C for 60 s, followed by 30 cycles of 95 °C for 15 s and 67 °C for 30 s. Gene expressions were calculated by the 2−ΔΔCT method. Expressions of fibronectin (FN), collagen IV (Col-IV), N-cadherin (N-cad), CDKN1B, and L13Rik were normalized by GAPDH, and expression of miR-2861 was normalized by U6. All primer sequences were listed in Table S1 .
Viplex fluor real time pcr system
Manufactured by Vivantis Technologies
Sourced in China, Malaysia
The ViPlex Fluor real-time PCR system is a laboratory instrument designed for the amplification and detection of nucleic acid sequences in real-time. It utilizes fluorescent technology to monitor the progress of the PCR reaction, allowing for quantitative analysis of target DNA or RNA.
Automatically generated - may contain errors
2 protocols using viplex fluor real time pcr system
1
Quantifying Gene Expression in Renal Tissues
2
Quantitative Gene Expression Analysis
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Total RNA in blood samples and mesangial cells was extracted with RNA simple Total RNA Kit (Tiangen, Beijing, China) according to the manufacturer's introduction. cDNA was synthesized with TIANScriptⅡRT Kit (Tiangen) at 42 ℃ for 60 min. qPCR was performed in triplicate with FastFire qPCR PreMix (SYBR Green) (Tiangen) on ViPlex Fluor Real-time PCR system (Vivantis, Selangor, Malaysia).
Thermocycling conditions were set as follows: 95 ℃ for 60 s, followed by 30 cycles of 95 ℃ for 15 s and 67℃ for 30s. Gene expressions were calculated by 2 -ΔΔCT method. Expressions of Fibronectin (FN), collagen IV (Col-IV), N-cadherin (N-cad), CDKN1B and L13Rik were normalized by GAPDH, and expression of miR-2861 was normalized by U6. All primer sequences were listed in Table-1.
Thermocycling conditions were set as follows: 95 ℃ for 60 s, followed by 30 cycles of 95 ℃ for 15 s and 67℃ for 30s. Gene expressions were calculated by 2 -ΔΔCT method. Expressions of Fibronectin (FN), collagen IV (Col-IV), N-cadherin (N-cad), CDKN1B and L13Rik were normalized by GAPDH, and expression of miR-2861 was normalized by U6. All primer sequences were listed in Table-1.
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