An alternative to culture-based cell death detection is an assessment of other cell viability indicators using fluorescent dyes, including membrane potential and membrane integrity. Live/Dead assays differentiates live and dead cells using membrane integrity as a proxy for cell viability and are based on a fluorescent staining procedure followed by detection using flow cytometry. However, samples preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.
An alternative to culture-based cell death detection is an assessment of other cell viability indicators using fluorescent dyes, including membrane potential and membrane integrity. Live/Dead assays differentiates live and dead cells using membrane integrity as a proxy for cell viability and are based on a fluorescent staining procedure followed by detection using flow cytometry. However, samples preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.
The challenge in isolating RNA from S. aureus cells is the disruption of the cell wall. A lot of protocols employ enzymatic digestion (pretreatment) which may affect gene expression patterns of certain genes. Therefore physical disruption using beads is considered to be the better alternative.
The biggest problem in isolating RNA from gram-positive bacteria is the disruption of the cell wall. A lot of protocols employ enzymatic digestion (pretreatment) which may affect gene expression patterns of certain genes. Therefore physical disruption using beads could be the best alternative.
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.
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.
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 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.
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.
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.
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