Hen eggs were purchased from Zhongbai Holdings Group Corporation (Wuhan, Hubei, China). Total plasma triglycerides (TG), total cholesterol (TC), HDL‐C, LDL‐C, glucose, alanine aminotransferase (ALT), aspartate aminotransferase (AST), C‐reactive protein (CRP), and urinary microalbumin kits were obtained from Daiichi Pure Chemicals. Plasma ghrelin kit was provided by Phoenix Pharmaceuticals. Plasma insulin kit was obtained from Mercodia AB and creatinine kit was obtained from Abcam. Total plasma apolipoprotein A‐I (apoA‐I) and apolipoprotein B (apoB) kits were obtained from Invitrogen. Ficoll‐Paque PREMIUM was obtained from GE Healthcare (1.077 g/ml). Ethylene diamine tetraacetic acid (EDTA) was obtained from Sigma‐Aldrich and TRIZOL reagent was obtained from Life Technologies corporation. The real‐time PCR Master Mix was provided by Toyobo and mRNA primers were obtained from Sango Biotech.
Mrna primers
Manufactured by Sangon
Sourced in China
MRNA primers are short, synthetic DNA sequences designed to specifically bind and initiate the amplification of mRNA targets. They serve as essential components for reverse transcription and quantitative PCR (RT-qPCR) studies.
Automatically generated - may contain errors
Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (5 mentions)
Mirna primer, by RiboBio (2 mentions)
Ethylenediaminetetraacetic acid edta, by Merck Group (1 mentions)
Ficoll paque premium, by GE Healthcare (1 mentions)
Realtime pcr master mix, by Toyobo (1 mentions)
Reverse transcript kit, by Takara Bio (1 mentions)
Sybr premix ex tag, by Takara Bio (1 mentions)
Viia 7 real time pcr system, by Thermo Fisher Scientific (1 mentions)
Kod sybr qpcr kit, by Toyobo (1 mentions)
5 protocols using mrna primers
1
Metabolic and Inflammatory Markers in Eggs
2
Quantitative RNA Expression Analysis
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Total cell RNA was extracted by using the TRIzol reagent (Invitrogen, CA, USA). Reverse transcription and the PCR process were performed according to the manufacturer’s instructions (Applied Biosystem, ThermoFisher Scientific, Shanghai, China). The mRNA primers were designed and synthesized by Sangon Biotech Co. Ltd. (Shanghai, China), and miRNA primers were synthesized by RiboBio Co. Ltd. (Guangzhou, China). The primer sequences are presented in Supplementary Table 1
3
Quantifying mRNA and miRNA Expressions
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Total RNA was extracted with TRIzol reagent (Invitrogen). cDNA was synthesized with a reverse transcript kit (Takara) according to the manufacturer’s instructions. qPCR was performed in a ViiA 7 real-time PCR system (Applied Biosystems) using the SYBR premix EX TAG (Takara). miRNAs were detected by using Bulge-loop primers designed from RiboBio Biotech (Guangzhou, China). mRNA primers were designed and synthesized from Sangon Biotech. β-actin and small nuclear RNA U6 were used as internal controls for mRNAs and miRNAs.
4
Comprehensive RNA Expression Analysis
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Total RNA was extracted using TRIzol reagent (Invitrogen; Thermo Fisher Scientific, Inc. Waltham, MA, USA). RT-qPCR was performed with the Toyobo RT kit and KOD SYBR® qPCR kit (Toyobo Life Science, Osaka, Japan) following the manufacturer's protocol. mRNA primers were synthesized by Sangon Biotech Co., Ltd. (Shanghai, China), and miRNA-specific RT primers and PCR primers were synthesized by Guangzhou RiboBio Co., Ltd. (Guangzhou, China). The PCR thermocycling conditions were as follows: 2 min at 98°C; 40 cycles of 10 sec at 98°C, 10 sec at 60°C, and 30 sec at 68°C; and 5 min at 72°C. Standard curve analysis was used to assess amplification efficiency. The primer sequences are listed in Table I . All mRNA expression was normalized to GAPDH, and miR-486-5p expression was normalized to U6. The expression levels of mRNAs and miRNAs were determined by the 2−ΔΔCq method. All experiments were performed in triplicate.
5
RNA Extraction and Reverse Transcription
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Total cell RNA was extracted by using Trizol reagent (Invitrogen, CA, USA). The reverse transcription and PCR process was performed according to the manufacturer’s instructions (Applied Biosystem, ThermoFisher Scientific, Shanghai, China). The whole procedure of the experiment was carried out according to our previous study [38 (link)]. The mRNA primers were designed and synthesized by the Sangon Biotech Co. Ltd. (Shanghai, China). And the miRNA primers were synthesized by RiboBio Co. Ltd (Guangzhou, China). The primer sequences were supplemented in Additional file 1 : Table S4.
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