rna-isolation-purification-tissue-rat-subcutaneous

Get tips on using Long Range DNA Ladder (linear scale) to perform DNA Ladder Long Range

Get tips on using Mouse RANKL ELISA Kit (TNFSF11) (ab100749) to perform ELISA Mouse - RANK L

Get tips on using TriDye™ Ultra Low Range DNA Ladder to perform DNA Ladder Low Range

Get tips on using Mouse TNFSF11/RANKL PicoKine™ ELISA Kit to perform ELISA Mouse - RANK L

Get tips on using E-Gel™ Ultra Low Range DNA Ladder to perform DNA Ladder Low Range

Get tips on using E-Gel™ Low Range Quantitative DNA Ladder to perform DNA Ladder Low Range

Get tips on using Mouse TRANCE/RANK L/TNFSF11 Quantikine ELISA Kit to perform ELISA Mouse - RANK L

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.

In ChIP, the most vital step is the binding of an antibody and choosing the right antibody. The binding affinity of different types of immunoglobulins to protein A or G differs significantly. Henceforth, it is recommended to choose either protein A or protein G coated beads. If you do not see any product in the positive control, add 5–10 μg of chromatin and 1–5 μg of antibody to each IP reaction and incubate with antibody overnight and an additional 2 hr after adding Protein G/A beads. If no product is observed in the experimental sample, add more DNA to the PCR reaction or increase the number of amplification cycles. Furthermore, if you have any problem with antibodies, make sure to use the ChIP-validated antibody.

DNA ladder is typically used as a reference to estimate the size of unknown DNA samples that are separated based on their mobility in an electrical field. The critical points for running a DNA ladder are compatibility with running buffer, agarose gel percentage, and choosing the correct range of DNA ladder for sizing DNA molecules.