DNA transfection Mammalian cells Immortalized cell lines

Get tips on using TRI Reagent® Sigma to perform RNA isolation / purification Cells - immortalized A2780ADR

Get tips on using TRI Reagent® Sigma to perform RNA isolation / purification Cells - immortalized CHO

Get tips on using Direct-zol RNA Kits to perform RNA isolation / purification Cells - immortalized DH82

Get tips on using miRCURY RNA Isolation Kit to perform RNA isolation / purification Cells - immortalized A549

Get tips on using miRCURY RNA Isolation Kit to perform RNA isolation / purification Cells - immortalized HeLa

Get tips on using EpiTect MSP Kit to perform PCR Methylation specific PCR - Mammalian DNA

Get tips on using QIAGEN Multiplex PCR Kit to perform PCR Multiplex PCR - Mammalian DNA

The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.

DNA isolation and purification is the first critical step in sample preparation that helps ensure optimal performance of downstream assays like PCR, microarrays, and sequencing. Failure in yielding high-quality DNA would be the major reason that DNA doesn't work for the downstream application. To circumvent this, one should follow the recommended storage conditions to minimize DNA degradation by nucleases and shouldn't overload the purification system.