Premier 5

Manufactured by Sangon

Sourced in China

Premier 5.0 is a laboratory equipment designed for general scientific applications. It provides core functionality for its intended use within a laboratory setting.

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Lab products found in correlation

Abi 3730xl dna analyzer, by Thermo Fisher Scientific (1 mentions) Rna pcr kit ver 3, by Takara Bio (1 mentions)

2 protocols using premier 5

Primer Design and Sequencing of GWAS Candidate Genes

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Premier 5.0 and Oligo 7.0 (Sangon Biotech Co., Ltd. Shanghai) were used to design primes for all exons and flanking regions (within 2 kb distance of 5′ and 3’ end) in three GWAS candidate genes. In total, 50 pairs of primers (Supplementary Table S1) were designed, including 16 for CACNB2, 18 for SLC39A12, and 16 for ZEB1. These primers were then synthesized at Sangon Biotech (Shanghai, China).
The DNA samples were extracted from frozen semen of 68 Holstein bulls (Beijing Dairy Cattle Center, Beijing, China) using a standard phenol-chloroform method, and then, DNA concentration were diluted to 200 ng/μl by adding TB buffer. We randomly mixed DNA samples into three pools (one pool contained samples from 28 bulls and two pools contained 20 samples each) and then used pooled samples for the subsequent PCR amplification [ABI3730XL DNA analyzer (Applied Biosystems, Foster, CA, United States)]. Sequencing data were aligned to the reference genome (UMD 3.1) using Ensemble (https://asia.ensembl.org/) and examined for potential polymorphism using Chromas (http://technelysium.com.au/wp/chromas/) and DNAman (https://www.lynnon.com/dnaman.html/). In total, 37 variants were detected from genes CACNB2, SLC39A12, and ZEB1.

Sammad A., Zhang H., Shi R., Dong Y., Luo H., Chen Z., Liu L., Guo G., Liu A, & Wang Y. (2022). A Post-GWAS Functional Analysis Confirming Effects of Three BTA13 Genes CACNB2, SLC39A12, and ZEB1 on Dairy Cattle Reproduction. Frontiers in Genetics, 13, 882951.

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Notch1 Gene Expression in Spinal Cord Tissue

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Total RNA in spinal cord tissue was extracted with Trizol and reverse transcription was conducted using the RNA PCR kit Ver.3.0 (TaKaRa, Dalian, Liaoning Province, China) according to the manufacturer's instructions. Primers were designed by Premier 5.0 (Sangon Biotech, Shanghai, China); Notch1 gene: forward primer 5′-GCA GCC ACA GAA CTT ACA AAT CCA G-3′, reverse primer 5′-TAA ATG CCT CTG GAA TGT GGG TGA T-3′ (689 bp); β-actin: forward primer 5′-GTG GGG CGC CCC AGG CAC CA-3′, reverse primer 5′-CTT CCT TAA TGT CAC GCA CGA TTT C-3′ (170 bp). Amplification conditions were as follows: denaturation at 94°C for 5 minutes, 29 cycles of 94°C for 30 seconds, 56°C for 40 seconds and 72°C for 50 seconds, followed by extension at 72°C for 10 minutes. Reactions were preserved at 4°C. DNA products were examined by 2% agarose gel electrophoresis. Target gene expression is presented as the ratio of optical density of the gene product versus the internal reference.

Guo Z.P., Huang M.N., Liu A.Q., Yuan Y.J., Zhao J.B, & Mei X.F. (2015). Buyang Huanwu decoction up-regulates Notch1 gene expression in injured spinal cord. Neural Regeneration Research, 10(8), 1321-1323.

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