Tianamp blood dna kit

As DNA from human or other non-human species may be mix in detected material, the TPI-plex amplification system was tested with DNA from a range of species including human, sheep, chick, duck, dog and rat, under the same PCR amplification condition to estimate potential interference. Genomic DNAs from human and dog samples were extracted from whole blood with TIANamp Blood DNA Kits^® (TIANGEN Biotech, Beijing, China), while others were extracted from fresh tissue (purchased from markets) using TIANamp Genomics DNA Kits^® (TIANGEN Biotech, Beijing, China). Each DNA was analyzed in duplicate and a negative control group was set up.

Genomic DNA samples were extracted from peripheral blood lymphocytes of patients using TIANamp Blood DNA kits (Tiangen Biotech CO., LTD. Beijing, China). DNA concentration and quality were assessed using absorbance spectrophotometry and agarose gel electrophoresis, respectively. The selection of IRS-1 tag SNP was performed with GEVALT 2.0 software (GEnotype Visualization and Algorithmic Tool) [16 (link), 17 (link)]. The SNP genotype data for China (CHB) population were downloaded from HapMap Project Browser, submitting a 100-kilobase pair region as a query, with a minor allele frequency cutoff of 0.05 and linkage disequilibrium (LD) measure r² threshold of 0.8. LD blocks were determined according to the gerbil algorithm [16 (link)]. Haplotype frequency was determined by means of the algorithms implemented in the GEVALT software, using 0.05 as the frequency threshold to define common haplotypes. The specific primers were designed by primer 5.0 software (see Additional file 1: Table S1). Genotypes were identified by polymerase chain reaction (PCR) and direct sequencing of genomic DNA.

Peripheral blood was collected from ulnar veins in the fasting and clearheaded state. Samples were centrifuged and serum was stored at – 80 °C until analysis. Genomic DNA was extracted utilizing TIANamp Blood DNA kits (TIANGEN, Beijing, China) in line with manufacturer’s instructions. VEGFR1 genotyping reactions were completed by Gene Company using KASP (Gene Company, Shanghai, China). Information of KASP primers (Primer_AlleleFAM, Primer_AlleleHEX and Primer_Common) was listed in Supplementary Table 2. To prove the reliability of genotyping results, five percent of the whole samples were repeatedly genotyped. Concordance rate of the repeated cases performed 100%, demonstrating that the results were reliable in this study.

Previous studies were systematically reviewed, and the National Center for Biotechnology Information database (NCBI; https://www.ncbi.nlm.nih.gov/) and Genome Reference Consortium Human Build 38 project (GRCh38; http://may2017.archive.ensembl.org/Homo_sapiens/Info/Index) were considered. Finally, seven GDF-15 gene polymorphisms (rs1055150, rs1058587, rs1059519, rs1059369, rs1227731, rs4808793, and rs16982345) were selected. All the loci ware according with pairwise tagging of HapMap population with r² ≥ 0.8, a minor allele frequency ≥5%, and Chinese Han Beijing (CHB) ethnicity. Genomic DNAs were extracted and purified from peripheral blood mononuclear cells by TIANamp Blood DNA kits (Tiangen, Beijing, China). Genotyping was conducted by the Kompetitive Allele-Specific PCR (KASP) method (Gene Company, Shanghai, China). Primer information for each polymorphism was listed in Supplementary Table 1.

Genomic DNA was extracted from venous blood samples using TIANamp Blood DNA kits (Catalog No. DP335-02; TIANGEN Biotech Co. Ltd., Beijing, China) following the manufacturer's instructions (17 (link)). NanoPhotometer® N50 (Implen GmbH., Munich, Germany) was used to assess DNA purity and concentration. Genomic DNA was amplified using the ABI-7900 real-time quantitative PCR instrument (Applied Biosystems, Foster City, CA, USA) and was subjected to HAAO rs3816183 TaqMan genotyping (18 (link)). PCR reactions were run as described in the previous study (19 (link)) using TaqMan® SNP Genotyping Assays (Catalog No: 4351379,C_180222_20, Thermo Fisher, USA) and TIANexact genotyping qPCR PreMix (Probe) (Catalog No. FP211-02; TIANGEN Biotech Co. Ltd., Beijing, China). In addition, 10% of DNA samples were selected randomly for second genotyping. The accuracy of data was ensured by the replicated samples with 100% consistency (19 (link)).

Genomic DNAs from whole blood were extracted by TIANamp Blood DNA Kits^® (TIANGEN Biotech, Beijing, China), following the manufacturer’s instructions. The hair shaft of each hair sample was cut off and the remaining part containing the hair follicle was placed into a 1.5 ml Eppendorf tube and washed with double distilled water and absolute ethyl alcohol, respectively and digested by proteinase K. The genomic DNAs from the hair samples were isolated using Chelex 100.