Immunohistochemistry Collagen Type I Goat Mouse

I have extracted RNA from brain tissue but my RNA concentrations are as low as 5ng/ml with my highest being around 80ng/ml. Will I be able to perform cDNA conversion with concentrations as low as these?

Get tips on using Human IGF1 ELISA Kit (ab100545) to perform ELISA Human - IGF-I

Get tips on using pDG101 to perform Protein Expression Prokaryotic cells - E. coli mScarlet-I

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.

Does anyone know how “strong” the PCR product of methylation specific PCR is? I kept my PCR products at 4C for about 3 weeks and then at room temperature for another week. Will I be able to use them for sequencing?

Get tips on using Rat IGF-1 ELISA Kit (ab213902) to perform ELISA Rat - IGF-I

Get tips on using Rat IGF-1 PicoKine™ ELISA Kit to perform ELISA Rat - IGF-I

Get tips on using Human IGF-1 PicoKine™ ELISA Kit to perform ELISA Human - IGF-I

HeLa cells I used are passage 40. Are these cells good to perform Cell cycle analysis on?

I have been doing multiplex PCR and qPCR separately. Is there a way to combine them in a single reaction using Sybr Green?