Bacterial cell culture media

Get tips on using Gibco™DMEM/F-12 to perform Stem cell Differentiation media mPSCs/mESCs differentiation to Embryoid body (EB)

Get tips on using Gibco™Glasgow's MEM (GMEM) to perform Stem cell Differentiation media mPSCs/mESCs differentiation to Embryoid body (EB)

Get tips on using Gibco™KnockOut™ DMEM to perform Stem cell Differentiation media mPSCs/mESCs differentiation to Embryoid body (EB)

Get tips on using STEMdiff™ Pancreatic Progenitor Kit to perform Stem cell Differentiation media Differentiation of Human hESCs into pancreatic progenitors

Get tips on using Gibco™KnockOut™ Serum Replacement to perform Stem cell Differentiation media hESCs differentiation into cortical neuroepithelium (NE)

Get tips on using Gibco™ PSC Cardiomyocyte Differentiation Kit to perform Stem cell Differentiation media hESCs or hiPSCs differentiation into Cardiomyocytes

Get tips on using STEMdiff™ Hematopoietic Kit to perform Stem cell Differentiation media hiPSCs differentiation into CD43+ primitive hematopoietic progenitor cells (HPCs)

Short hairpin or small hairpin RNA (shRNA) is artificial RNA, which has a hairpin loop structure, and uses inherent microRNA (miRNA) machinery to silence target gene expression. This is called RNA interference (RNAi). These can be delivered via plasmids or viral/bacterial vectors. Challenges in shRNA-mediated gene silencing include: 1. Off-target silencing, 2. Packaging shRNA encoding lentivirus, and 3. Stable transduction in cells. RNAi have been designed to have anywhere from 19-27 bs, but the most effective design has 19 bp. In case commercial shRNAs are not available, potential target sites can be chosen within exon, 5’- or 3’ UTR, depending on which splice variants of the gene are desired. One should use the latest algorithms and choose at least two different sequences, targeting different regions, in order to have confidence in overcoming off-target effects. A BLAST search after selecting potential design will eliminate potential off-target sequences. For the second challenge, sequencing the vector using primers for either strand (50-100 bp upstream) is suggested, along with using enzymatic digestion on agarose gel for the vector. Next, once the shRNA-containing vector is packaged in a virus, it is important to check the viral titer before transduction. Finally, using a marker in the lentiviral vector (fluorescent protein or antibiotic resistance), along with qPCR for target gene expression can help in determining efficacy of transduction and shRNA on its target site.

Get tips on using IMAGEN™ Respiratory Virus Screen Kit using Direct Immunofluorescence Assay to perform Cell Culture Contamination Detection Kit Virus

Get tips on using IMAGEN™ Parainfluenza Virus Group Kit using Direct Immunofluorescence Assay to perform Cell Culture Contamination Detection Kit Virus