7500 system sds v 1

Fourteen tissues were obtained from three adult Qinchuan cattle. Total RNA was extracted from the tissues using a Total RNA kit (Tiangen, Beijing, China) and then reverse-transcribed using a PrimeScript™ RT reagent Kit with gDNA Eraser (Perfect Real Time) (TaKaRa, Dalian, China). cDNA from reverse transcription of individual animal samples for each tissue was pooled for qPCR which was performed using a SYBR Green PCR Master Mix kit (TaKaRa, Dalian, China) and 7500 System SDS V 1.4.0 (Applied Biosystems, USA). All of the primers used in the real-time PCR experiment are listed in Supplementary Table S1. The data were normalized against those obtained for the housekeeping gene, β-actin (ACTB), which was used as an endogenous control gene. The relative expression levels of the target mRNAs were calculated using the 2^−ΔΔCt method52 (link).

Fifteen tissues (heart, liver, spleen, kidney, rumen, reticulum, omasum, abomasa, small intestine, large intestine, subcutaneous fat, longissimus thoracis, soleus, psoas and testicle) were obtained from three adult Qinchuan cattle. The longissimus thoracis samples were collected during eight developmental stages from male Qinchuan cattle, including 1, 3, 6, 12, 18, 24, 36 and 48 months after birth, and three parallel individuals were sampled during each period. Total RNA was extracted from the tissues using a Total RNA Kit (Tiangen, Beijing, China) and then reverse-transcribed using a PrimeScript™ RT Reagent Kit (Perfect Real Time) (TaKaRa, Dalian, China). The reaction was performed using a SYBR Green PCR Master Mix Kit (TaKaRa) on a 7500 System SDS V 1.4.0 (Applied Biosystems, USA). All primers used in the real-time PCR experiments are listed in Supplementary Table S1. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was invoked as the endogenous control gene. The relative expression levels of the target mRNAs were calculated using the 2^−ΔΔCt method^{43 (link)}.

Three adult Qinchuan cattle served as a source of 14 tissue samples, from which total RNA was isolated using the Total RNA kit (Tiangen, Beijing, China). cDNA was generated through use of the PrimeScript™ RT reagent Kit (TaKaRa, Dalian, China), and individual animal sample tissue cDNA were pooled before performing a qPCR analysis using a SYBR Green PCR Master Mix kit (TaKaRa) with a 7500 System SDS V 1.4.0 (Applied Biosystems, Foster City, CA, USA). Table S1 (Supplementary Material) contains the primers used in the present study. All gene expressions were normalized with of β-actin (ACTB) expression, and the 2^−ΔΔCt method was used to compare gene expressions [21 (link)].