Site Directed Mutagenesis (SDM) Human Point mutation LNCaP

Cells are sourced from various tissues to grow them in in-vitro conditions. Therefore, cell specific nutrients are important for their survival, maintenance and growth. Determining the appropriate cell culture media is a challenge if you are growing a cell line or a microorganism for the first time. Established cell lines, primary cells, stem cells, bacteria and Yeast all require varied nutrients from basic to complex. Based on the cell type, one can easy find what media and nutrients your peers have used before you try to reinvent the wheel.

When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel.

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.

Microarrays enable researchers to monitor the expression of thousands of genes simultaneously. However, the sensitivity, accuracy, specificity, and reproducibility are major challenges for this technology. Cross-hybridization, combination with splice variants, is a prime source for the discrepancies in differential gene expression calls among various microarray platforms. Removing (either from production or downstream bioinformatic analysis) and/or redesigning the microarray probes prone to cross-hybridization is a reasonable strategy to increase the hybridization specificity and hence, the accuracy of the microarray measurements.

Get tips on using ON-TARGETplus Human MDK (4192) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - LNCap MDK

Get tips on using ON-TARGETplus Human MID1 (4281) siRNA - Individual to perform siRNA / miRNA gene silencing Human - LNCap MID1

Get tips on using ON-TARGETplus Human AR (367) siRNA - Individual to perform siRNA / miRNA gene silencing Human - LNCap AR

Get tips on using STEAP1 siRNA to perform siRNA / miRNA gene silencing Human - LNCap STEAP1

Get tips on using siRNA SKIL to perform siRNA / miRNA gene silencing Human - LNCap SKIL

Get tips on using siRNA PIN1 to perform siRNA / miRNA gene silencing Human - LNCap PIN1