siRNA / miRNA gene silencing Rat MTLn3 (rat mammary adenocarcinoma breast cancer cell line)

Get tips on using Anti Type X Collagen (raised against rat) pAb (Rabbit, Antiserum) to perform Immunohistochemistry Mouse - Col X

Get tips on using A2B5 Antibody, anti-human/mouse/rat, APC to perform Flow cytometry Anti-bodies Human - A2B5

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.

The process of RNA extraction from bacteria, in general, involves an RNA-protective, effective lysis of bacterial cell wall (which may pose difficulties). EDTA promotes loss of outer membrane to provide lysozyme with access to peptidoglycan. Another common method for cell wall lysis is mechanical disruption using a homogenizer (applied for gram-positive bacteria and some strains of gram-negative bacteria). Following lysis, it is necessary to disrupt protein-nucleic acid interactions, which can be achieved by adding sodium dodecyl sulfate (SDS). Next step involves using phenol-chloroform-isoamyl alcohol extraction, where RNA can be obtained from the bottom organic phase, the top phase consists of DNA and the interphase contains proteins. Isoamyl alcohol is an inert and optional addition to this mixture and is added as an anti-foaming reagent to reduce the interphase. Following RNA extraction, the samples should be checked for its quality by gel electrophoresis (23S and 16S rRNAs and 5s rRNA and tRNA bands) or UV spectrophotometric or fluorescence methods.

Get tips on using Biotin Rat Anti-Mouse OX40 Ligand (CD252) to perform Flow cytometry Anti-bodies Mouse - CD252/OX40L

Get tips on using PE-CF594 Rat Anti-Mouse Siglec-F to perform Flow cytometry Anti-bodies Mouse - Siglec F

Get tips on using ON-TARGETplus Human RAD51 (5888) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - MDA-MB-231 RAD51

Get tips on using pSUPER.retro.puro to perform shRNA gene silencing Rat - MM1 SSH1