background: breast cancer is one of the most prevalent malignancies among women in various countries. her2 overexpression, which is due to different reasons, occurs in 20-30% of breast cancers. her2 gene encodes an 185kda transmembrane glycoprotein with 1255 amino acids. this active product triggers downstream intracellular signaling pathways inducing cell proliferation and cell survival. these...

Small interfering RNA (siRNA)-induced gene silencing shows great promise in genomic research and therapeutic applications. siRNA duplexes are typically assembled from complementary synthetic oligonucleotides. High-purity single-stranded species are required for in vivo applications. Methods for separation, characterization, and purification of short RNA strands have been developed based on reve...

Norgen Biotek has developed various products (Plasma/Serum Circulating RNA Purification Kits and Exosome RNA Purification Kits) that are specifically optimized for purifying circulating or exosomal RNA (large RNA and microRNA) from plasma/serum or urine with the following advantages: 1) Simple and Fast Procedure with no lengthy phenol:chloroform extraction, filtration or precipitation 2) Scalab...

RNA-centric biochemical purification is a general approach for studying the functions and mechanisms of noncoding RNAs. Here, we describe the experimental procedures for RNA antisense purification (RAP), a method for selective purification of endogenous RNA complexes from cell extracts that enables mapping of RNA interactions with chromatin. In RAP, the user cross-links cells to fix endogenous ...

An improved purification procedure for Thermotoga maritima RNA-polymerase holoenzyme was developed. The enzyme is highly active with poly dAT or T7 phage DNA as template. DNA gyrase was found to be a side product of this RNA-polymerase purification. The genes for the large subunits beta and beta' of RNA-polymerase were cloned and sequenced. The phylogenetic position of T.maritima within the bac...

The chemical synthesis of backbone deuterium labelled [r(CGCGAAU*U*CGCG)]2 (U* = [5'-2H]U) is described. An efficient purification procedure was developed using a polymeric reverse phase (PRP) HPLC column at 60 degrees C. This procedure provided pure RNA dodecamer in the multi-milligram quantities (39% overall yield) necessary for dynamics studies using solid-state deuterium NMR. The purificati...

Although RNA-based biological processes and therapeutics have gained increasing interest, purification of in vitro transcribed RNA generally relies on gel-based methods that are time-consuming, tedious and denature the RNA. Here, we present a reliable procedure for affinity batch purification of RNA, which exploits the high-affinity interaction between the boxB RNA and the N-peptide from bacter...

Abstract Using RNA antisense purification and mass spectrometry, we identified more than 100 human proteins that directly specifically bind SARS-CoV-2 in infected cells. To gain insights into the functions of selected interactors, applied genetic perturbation pharmacological inhibition experiments, mapped contact sites on viral RNA. This led to identification host dependency factors defense str...