We re-selected 6 fast- and slow-growing Jinghai yellow chickens (3 slow-growing chickens and 3 fast-growing chickens) to collected leg muscles. Total RNAs were isolated with Trizol (TIANGEN, Beijing, China) and miRNAs were reverse transcribed into cDNA with the miRNA First-Strand cDNA kit (TIANGEN, Beijing, China). The forward primers used for quantification in the study were designed using Primer 5.0 and U6 small nuclear RNA (U6) was used for the housekeeping gene. The qPCR was conducted on QuantStudio 3 with the miRNA qPCR kit (TIANGEN, Beijing, China), and the reverse primers also came from the qPCR kit. The relative expression of miRNAs was calculated using the 2−ΔΔCT method (Rao et al., 2013 (link)).
Quantstudio 3
Manufactured by Tiangen Biotech
Sourced in China
The QuantStudio 3 is a real-time PCR system designed for gene expression analysis, genotyping, and other quantitative PCR applications. It features a 96-well block format and supports a variety of chemistries and fluorescent dyes. The system provides accurate and precise data with intuitive software for data analysis and reporting.
Automatically generated - may contain errors
2 protocols using quantstudio 3
1
Differential miRNA Expression in Chicken Growth
2
Quantifying PCSK9 mRNA Knockdown in Liver
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TRIzol (TIANGEN Biotech, China) was used to extract RNA from liver tissue, and a FastKing RT kit (with gDNase) (TIANGEN Biotech, China) was used to reverse transcribe the RNA. Thermo Fisher Scientific QuantStudio3 was used to perform quantitative PCR (qPCR) on liver tissue cDNA using SuperReal PreMix Plus (TIANGEN). AAV plasmid expression was measured, and Pcsk9 mRNA knockout levels confirmed (Fig. S11 ). Liver tissues were fixed with 4% formalin overnight before being embedded in paraffin, sectioned, and stained with hematoxylin and eosin (H&E)46 (link). The liver sections were prepared by Servicebio (https://www.servicebio.cn/ ).
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