Tiangen dna extraction kit

Whole peripheral blood (PB) was collected from each subject. Mononuclear cells were separated by using Ficoll-Paque (Sigma, Saint Louis, MO, USA) density gradient centrifugation. Genomic DNA was extracted from PB mononuclear cell using the Tiangen DNA extraction kit (Tiangen Biotech Co. Ltd, Beijing, China) according to the recommended procedure. The concentration and purity of DNA product were determined by its optical absorbance and were diluted to 50 ng/μL.

In total, 229 Hulun Buir lambs (male = 106, female = 123), which were born in March 2019 on Hulun Buir sheep farms (Hulun Buir city, Inner Mongolia, China), were investigated. The animals were grazed in identical conditions. The birth weight (BW), weaning weight adjusted at 4-month-old (WW) and body weight at 9-month-old (NBW) were recorded. Meanwhile, average daily gains (ADG) during birth to weaning, weaning to 9-month-old and birth to 9-month-old periods were calculated. Body height (BH), body length (BL) and chest girth (CG) were measured at weaning and at 9 months of age, respectively. Approximately 1 cm³ marginal ear tissues were collected and preserved in 95% ethanol. Genomic DNA of Hulun Buir sheep was extracted using a Tiangen DNA extraction kit (Tiangen Biotech Co., Ltd, Beijing, China) and stored at −20 °C for PCR amplification.

Genomic DNA was extracted from peripheral blood samples (2–3 mL) of patients and their parents using the Tiangen DNA extraction kit (Tiangen Biotech, China). DNA quality and quantification were assessed using a NanoDrop 2000 (Thermo Fisher, USA).

DNA was extracted from umbilical cord tissues (about 5 cm from placenta attachment) of three infants with single chorionic triplets and preterm live birth after single blastocyst transplantation and double blastocyst transplantation double chorionic twins and two full-term live births using the Tiangen DNA Extraction Kit (TIANGEN, Beijing, China) according to manufacturer's instructions. DNA was sent directly to Gene Tech (Shanghai) Co, Ltd. (Gene Tech, Shanghai, China) for pyrosequencing to detect the methylation level of CpG island in CGRP promoter region. The sequencing primers, conditions, and sequencing sequences used are shown in Figure 1.

Blood samples (1–2 mL) were collected from the probands and their parents. Genomic DNA was extracted using a Tiangen DNA extraction kit (Tiangen Biotech, Beijing, China) according to the manufacturer’s instructions and quantified spectrophotometrically by NanoDrop 2000 manufacturer (ThermoScientific, USA).

Total RNA from MDA-MB-231 and BT-549 cells was isolated using TRIzol reagent (Invitrogen, Carlsbad, CA, USA). The genomic DNA (gDNA) from BC cells was extracted by a TIANGEN DNA extraction kit (#DP304, TIANGEN, Beijing, China). For the actinomycin D assay, BC cells were treated with actinomycin D prior to RNA extraction. For the RNase R assay, total cell RNA (2 μg) was digested with 3 U/μg RNase R reagent (#RNR07250, Biosearch Technologies, Petaluma, CA, USA) at 37 °C for 15 min and 85 °C for 3 min. Following treatment with RNase R and actinomycin D, circRNF10 expression and linear RNF10 were investigated using qRT-PCR.

Genomic DNA was extracted from peripheral blood samples using a TIANGEN DNA extraction kit (TIANGEN, Beijing, China). Sample preparation and pretreatment for next generation sequencing were performed using the SureSelect Human All Exon V5 kit (Agilent). Exome sequencing was carried out on an HiSeq2500 system (Illumina).