The following tests were performed on the control and patient groups: All abnormal results on HPLC were confirmed with alkaline and acid pH Hb electrophoresis (HYDRASYS system, Sebia-Norcross; SEBIA-INC, USA) 4. Multiplex PCR was performed as described by Chong et al. [3] DNA was extracted using a High Pure PCR preparation kit (Roche Diagnostics, Germany).
High pure pcr preparation kit
Manufactured by Roche
Sourced in Germany
The High Pure PCR preparation kit is a lab equipment product designed for the purification of PCR-amplified nucleic acids. It facilitates the extraction and purification of DNA or RNA from various sample types. The kit provides the necessary components and protocols to prepare purified nucleic acid samples suitable for downstream applications such as PCR, sequencing, or other analysis methods.
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Lab products found in correlation
2 protocols using high pure pcr preparation kit
1
Confirmatory Hb Electrophoresis and PCR Analysis
2
Methylation Analysis of Genomic DNA
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DNA was extracted from whole blood using the High Pure PCR Preparation Kit (Roche), and then, it was sonicated to obtain size range of fragment (200-1000 bp) optimal for MeDIP analysis.
Sonicated DNA was subjected to MeDIP process in which all the methylated DNA sequences are pulled down by a monoclonal antibody against 5-methylcytosine. MeDIP was performed using the Methylated DNA Capture Kit (Epigentek) according to the manufacturer's instructions. Enriched methylated DNA sequences were amplified in quantitative real-time polymerase chain reaction (qPCR) using primers specific to H19 gene as the endogenous positive control (hemimethylated gene) and GAPDH as the unmethylated gene. The same amplification reaction was done on input DNA samples which were the fraction of sonicated DNA including both methylated and unmethylated sequences that have been left before MeDIP analysis to be used as the reference DNA control. Enrichment was calculated using the following equation 20
Sonicated DNA was subjected to MeDIP process in which all the methylated DNA sequences are pulled down by a monoclonal antibody against 5-methylcytosine. MeDIP was performed using the Methylated DNA Capture Kit (Epigentek) according to the manufacturer's instructions. Enriched methylated DNA sequences were amplified in quantitative real-time polymerase chain reaction (qPCR) using primers specific to H19 gene as the endogenous positive control (hemimethylated gene) and GAPDH as the unmethylated gene. The same amplification reaction was done on input DNA samples which were the fraction of sonicated DNA including both methylated and unmethylated sequences that have been left before MeDIP analysis to be used as the reference DNA control. Enrichment was calculated using the following equation 20
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