Genomic DNA was isolated from each patient’s peripheral blood lymphocytes using BioTek DNA Whole-blood Mini Kit (BioTek, Beijing, China) or from saliva using Oragene tubes (DNA Genotek, Canada). Whole exome sequencing (WES) was performed by the Beijing Genomic Institute (BGI, Beijing, China) using DNA from lymphocytes from 6 non-consanguineous individuals with tooth agenesis. The coding exons of the BMP4 gene of all patients were amplified by polymerase chain reaction (PCR) using the following primer sequences: exon 3-F (5′-CCATCTTGCCCCTCCATTTCTA-3′); exon 3-R (5′-CTTCTTCCCCAGGGCTTTCACT-3′); exon 4a-F (5′-TGCTTATTTTCCCCCAGTAGGT-3′); exon 4a-R (5′-CCCGCTGTGAGTGATGCTT-3′); exon 4b-F (5′-GGGCCAGCATGTCAGGATTAGC-3′) and exon 4b-R (5′-GTGGGTGAGTGGATGGGAACG-3′). The amplified PCR products were sequenced by Tsingke Biological (Beijing, China) and the results were blasted on NCBI.
Biotek dna whole blood mini kit
Manufactured by Agilent Technologies
Sourced in China
The BioTek DNA Whole-blood Mini Kit is a laboratory equipment product designed for the extraction and purification of genomic DNA from whole blood samples. It provides a simple and efficient method to obtain high-quality DNA for various downstream applications.
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Lab products found in correlation
2 protocols using biotek dna whole blood mini kit
1
Exome Sequencing of Tooth Agenesis
2
Whole Exome Sequencing for Orodental Disorder
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Peripheral blood genomic DNA was extracted using the BioTek DNA Whole-Blood Mini Kit (BioTek, Beijing, China), referring to the instructions. WES was performed in all the five available family members by Beijing Angen Gene Medicine Technology (Beijing, China) using the Illumina-X10 platform to identify the potential pathogenic gene variants. Based on the WES results, orodental-related genes were annotated [23 (link)]. Then, we filtered all the nonsynonymous single-nucleotide variants and insertions/deletions according to the MAF ≤ 0.01 in the databases, including the single-nucleotide polymorphism database (dbSNP, http://www.ncbi.nlm.nih.gov/projects/SNP/snp_summary.cgi , accessed on 1 October 2020) and the genome aggregation database (gnomAD, http://gnomad.broadinstitute.org accessed on 1 October 2020). The variants that affected protein function were predicted based on the results obtained from ReVe [24 (link)], rare-exome-variant ensemble learner (REVEL) [25 (link)], and combined annotation-dependent depletion (CADD) [26 (link)].
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