The RNA-guided CRISPR-Cas9 nuclease system has revolutionized the genome editing practices. For the most part, the Cas9-mediated genome editing is performed either via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, However, designing of specific sgRNAs and minimizing off-target cleavage mediated mutagenesis are the major challenges in CRISPR-Cas based genome editing. To circumvent these issues, we can take advantages of many available tools and approaches for sgRNA construction and delivery.
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.
Get tips on using Glut1 siRNA and shRNA Plasmids (h) to perform RNA sequencing Human - HT-1376 (urinary bladder cell line)
Get tips on using CD74 siRNA and shRNA Plasmids (h) to perform RNA sequencing Human - HT-1376 (urinary bladder cell line)
Get tips on using Glut1 siRNA and shRNA Plasmids (h) to perform siRNA / RNAi /miRNA transfection Human Cells - HT-1376 GLUT1
Get tips on using CD74 siRNA and shRNA Plasmids (h) to perform siRNA / RNAi /miRNA transfection Human Cells - HT-1376 CD74
Get tips on using gRNA_Cloning Vector to perform CRISPR Mouse - Deletion RMA cells Trh4
I intend to use iScript cDNA Synthesis Kit in order to synthesize cDNA for qPCR. I have confirmed that my RNA is pure however, according to my extraction protocol I have suspended the RNA in TE buffer containing 1mM EDTA. Will the presence of EDTA have an effect on cDNA synthesis?
Get tips on using MessageAmp II aRNA Amplification Kit to perform Microarray RNA amplification & Labeling - Rhesus monkey brain tissue Biotin
Get tips on using MessageAmp II aRNA Amplification Kit to perform Microarray RNA amplification & Labeling - Human brain tissue Cyanine 3
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