Expression levels of miRNA were determined using quantitative real-time quantitative PCR with The All-in-OneTM miRNA qRT-PCR Detection Kit (GeneCopoeia, USA) and mRNA qRT-PCR detection kit (GeneCopoeia, USA) was used to determine mRNA expression level in real-time quantitative PCR. Real-time quantitative PCR analysis was performed on a Bio-Rad iCycler system (Bio-Rad, Hercules, CA). Rno-U6 served as a loading control for the sample to test for miRNA and GAPDH served as a loading control for the sample to test for mRNA, mRNA expression levels and miRNA expression levels were evaluated using the 2 − (ΔΔCt) method. Sequences of specific primers are listed in Additional file 1 : Table S1.
Mrna qrt pcr detection kit
Manufactured by GeneCopoeia
Sourced in United States
The mRNA qRT-PCR detection kit is a laboratory equipment designed to quantify the expression levels of specific mRNA molecules using real-time reverse transcription-polymerase chain reaction (qRT-PCR) technology. The kit provides the necessary reagents and components to perform accurate and sensitive mRNA quantification.
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Lab products found in correlation
3 protocols using mrna qrt pcr detection kit
1
Quantifying miRNA and mRNA Expression
2
RNA Extraction and qRT-PCR Analysis
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Total RNA was extracted from corpus cavernosum tissue using the AxyPrep Multisource Total RNA Miniprep Kit (Axygen Scientific, Tewksbury, MA, USA), while cDNA was reverse transcribed using the SureScript™ First-Strand cDNA Synthesis Kit (GeneCopoeia, Rockville, MD, USA). qRT-PCR was conducted with a Bio-Rad iCycler system (Bio-Rad) using an mRNA qRT-PCR detection kit (GeneCopoeia). The program included 40 cycles (95°C, 10 s; 56°C, 30 s; and 72°C, 30 s). GAPDH was used as the loading control to detect mRNA, and mRNA expression levels were determined with the 2−ΔΔCt method. The primer sequences used are contained in Table 1
3
Hippocampal DG mRNA Expression Analysis
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According to the manufacturers’ instructions, total RNA was isolated from hippocampal DG regions using the Trizol kit (Invitrogen) and was reverse transcribed into cDNA using reverse transcriptase (GeneCopoeia, USA). Quantitative real-time expression levels of mRNA were determined using the mRNA qRT-PCR detection kit (GeneCopoeia, USA). Real-time quantitative PCR analysis was performed on a Bio-Rad iCycler system (Bio-Rad, Hercules, CA). GAPDH served as a loading control for the samples to test for mRNA, with mRNA expression levels evaluated using the 2 − (ΔΔCt) method. Sequences of specific primers are listed in Additional file 4 : Table S1.
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