cell-cycle-assay-human-hela

Get tips on using Guava Cell Cycle Reagent for Flow Cytometry to perform Cell cycle assay human - Jurkat

Get tips on using Guava Cell Cycle Reagent for Flow Cytometry to perform Cell cycle assay human - K562

Get tips on using Guava Cell Cycle Reagent for Flow Cytometry to perform Cell cycle assay human - SKOV3

Get tips on using Guava Cell Cycle Reagent for Flow Cytometry to perform Cell cycle assay human - OVCAR-5

Get tips on using Guava Cell Cycle Reagent for Flow Cytometry to perform Cell cycle assay human - THP-1

Get tips on using Guava Cell Cycle Reagent for Flow Cytometry to perform Cell cycle assay human - MCF-7

Get tips on using Guava Cell Cycle Reagent for Flow Cytometry to perform Cell cycle assay human - SH-SY5Y

Though DNA quantification is but one small step in the multifaceted DNA sample preparation workflow, it can have large implications on the performance and validity of conclusions drawn from downstream assays. Major challenges include accuracy, precision, reproducibility, and detection of present contamination. Among UV spectrophotometry, fluorescence and real-time PCR based methods, the quantification method should be chosen based on the requirement of the downstream assay.

DNA-protein interactions are studied by using ChIP. The basic steps in this technique are crosslinking, sonication, immunoprecipitation, and analysis of the immunoprecipitated DNA. During ChIP, if chromatin is under-fragmented or fragments are too large which can lead to the increased background and lower resolution. Shorter cross-linking times (5-10 min) and/or lower formaldehyde concentrations (<1%) may improve shearing efficiency. If Chromatin is over-fragmented, then optimize shearing conditions for each cell type to improve ChIP efficiency. Over-sonication of chromatin may disrupt chromatin integrity and denature antibody epitopes. If you do not see any product or very little product in the input PCR reactions, add 5–10 μg chromatin per IP.

Cell Invasion or Cell Migration assays are technically challenging to set up as the gradient between the two compartments equilibrates in time during the assay. It is also problematic to view cells and for cells to migrate through a non-physiologic polycarbonate or polypropylene filter. Care must be taken while loading the well with cells to form a single cell suspension. Precaution must be taken while trypsinization (under-trypsinization can lead to cell clumping while over-trypsinization could strip off adhesion molecules necessary for migration). This leads to difficulty in getting significant results, when only small numbers of cells cross the filter or when the distribution and/or staining of the cells is uneven.