Genomic DNA was extracted from peripheral blood samples using the Maxwell blood DNA purification kit (Promega Corporation, Madison, WI) following standard protocols. Subsequently, mutation analysis was performed by Sanger sequencing of PCR products using primers specific to each CNGB3 (NM_019098.4 ; hg19) region (Table 1 ). Segregation analysis was performed on all families in which we were able to recruit additional family members.
Maxwell blood dna purification kit
Manufactured by Promega
Sourced in United States
The Maxwell Blood DNA Purification Kit is a laboratory instrument designed to efficiently extract and purify DNA from whole blood samples. It utilizes magnetic bead technology to capture and isolate DNA, providing a reliable and standardized method for DNA extraction.
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5 protocols using maxwell blood dna purification kit
1
Genomic DNA Extraction and Mutation Analysis in CNGB3
2
ALDH2 Genotyping from Blood DNA
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Details of the DNA extraction and genotyping procedures have been previously described16
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21)
. Genomic DNA was extracted from peripheral blood using the Maxwell Blood DNA purification Kit (Promega Corporation, Madison, WI). The SNP rs671 in ALDH2 was analyzed via the TaqMan probe assay (Applied Biosystems, Waltham, MA). To determine the ALDH2-rs671 genotypes, a commercially available primer and probe were purchased from the Assay-on-Demand system (C_11703892_10). Fluorescence of polymerase chain reaction products was measured using the ABI PRISM 7900HT sequence detector (Applied Biosystems). In all samples, the genotype obtained by sequencing coincided with that analyzed by the TaqMan probe assay.
,
21)
. Genomic DNA was extracted from peripheral blood using the Maxwell Blood DNA purification Kit (Promega Corporation, Madison, WI). The SNP rs671 in ALDH2 was analyzed via the TaqMan probe assay (Applied Biosystems, Waltham, MA). To determine the ALDH2-rs671 genotypes, a commercially available primer and probe were purchased from the Assay-on-Demand system (C_11703892_10). Fluorescence of polymerase chain reaction products was measured using the ABI PRISM 7900HT sequence detector (Applied Biosystems). In all samples, the genotype obtained by sequencing coincided with that analyzed by the TaqMan probe assay.
3
Automated DNA Isolation from Peripheral Blood
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Peripheral blood samples (5 mL) were collected in Vacutainer vacuum blood tubes with EDTA and then subjected to a specific lysis pretreatment before proceeding to the DNA isolation methods. Pretreatment consisted of mixing 1:1 blood and denaturing lysis buffer (6M guanidine hydrochloride, 10 mM urea, 10 mM TRIS-HCL, 20% (v/v) Triton X-100; pH was adjusted to 4.4) through brief shaking. The samples were incubated at 70°C for 10 min. Before proceeding to the next steps, the samples were kept for at least 48 h at room temperature.
The automated purification protocol of the Maxwell Blood DNA Purification kit (Promega Biotech) was chosen according to the guidelines approved by HUyP-La Fe and was used following the manufacturer’s instructions. The initial volume for each of the blood sample was 400 μL of pretreated peripheral blood.
The automated purification protocol of the Maxwell Blood DNA Purification kit (Promega Biotech) was chosen according to the guidelines approved by HUyP-La Fe and was used following the manufacturer’s instructions. The initial volume for each of the blood sample was 400 μL of pretreated peripheral blood.
4
Illumina PsychArray Genotyping Protocol
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Genomic DNA was extracted from blood samples with the Maxwell Blood DNA purification kit (Promega, Madison, WI, USA) in accordance with the manufacturer’s protocol and the samples were genotyped using the Illumina Infinium PsychArrayBeadChip (Illumina, San Diego, CA, USA).
5
Blood DNA Extraction and Genotyping
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Genomic DNA was extracted from blood samples using the Maxwell Blood DNA purification kit (Promega, Madison, WI, USA) in accordance with the manufacturer's protocol and genotyped using the Illumina Infinium PsychArrayBeadChip (Illumina, San Diego, CA, USA).
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