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qRT-PCR method

WY Wei Yuan
HQ Hai Qin
HB Huan Bi
DZ Depeng Zhao
YZ Yiyu Zhang
WC Wei Chen
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After 48 h transfection period, total RNA was extracted from cells using the Trizol method, followed by cDNA synthesis using HiFiScript cDNA Synthesis Kit (Purdy Bio, Shanghai, China). SYBR Green kit (Tiangen Bio, Beijing, China) was utilized to detect miRNA expression levels, and 2×Taq MasterMix (Kangwei Century Bio, Beijing, China) was used to detect mRNA expression levels, according to the kit instructions. The qRT-PCR reaction system and reaction conditions varied (Supplementary Table 6). The qRT-PCR technique was employed to determine the changes in mRNA expression levels of skin color-related genes in Jianbai Xiang pig melanocytes transfected with pEGFP-N3-TYRP1 and TYRP1-siRNA. The ssc-miR-221-3p, TYR, TYRP1, DCT, and 5 S rRNA internal reference primer sequences were shown in Supplementary Table S7.

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