Search results
Results From The WOW.Com Content Network
Disassociate the RNA from the protein of interest. Isolate the RNA fragments from the protein using a centrifuge. Use Reverse Transcription PCR to convert the RNA fragments into cDNA (DNA that is complementary to the RNA fragments). Fluorescently label these cDNA fragments. Prepare the gene chip. This is a small chip that has DNA sequences ...
Immunoprecipitation of intact protein complexes (i.e. antigen along with any proteins or ligands that are bound to it) is known as co-immunoprecipitation (Co-IP). Co-IP works by selecting an antibody that targets a known protein that is believed to be a member of a larger complex of proteins.
Cross-linking and immunoprecipitation (CLIP, or CLIP-seq) is a method used in molecular biology that combines UV crosslinking with immunoprecipitation in order to identify RNA binding sites of proteins on a transcriptome-wide scale, thereby increasing our understanding of post-transcriptional regulatory networks.
RNA partitions in the aqueous phase, while proteins and DNA partition into the organic/interphase (left). The RNA is then precipitated in an alcohol (right). Acid guanidinium thiocyanate-phenol-chloroform extraction (abbreviated AGPC) is a liquid–liquid extraction technique in biochemistry and molecular biology.
The RNA is then reverse transcribed, causing most cDNAs to truncate at the crosslink site, and the key innovation and unique feature in the development of iCLIP was to enable such truncated cDNAs to be PCR amplified and sequenced using a next-generation sequencing platform. iCLIP also added a random sequence (unique molecular identifier, UMI ...
Fragmented gDNA is incubated with the DNA-RNA structure-specific S9.6 mAb. This step is unique for the DRIP-seq protocol, since it entirely relies on the high specificity and affinity of the S9.6 mAb for DNA-RNA hybrids. The antibody will recognize and bind these regions dispersed across the genome and will be used for immunoprecipitation.
In chemistry, coprecipitation (CPT) or co-precipitation is the carrying down by a precipitate of substances normally soluble under the conditions employed. [1] Analogously, in medicine, coprecipitation (referred to as immunoprecipitation) is specifically "an assay designed to purify a single antigen from a complex mixture using a specific antibody attached to a beaded support".
Ethanol precipitation is a method used to purify and/or concentrate RNA, DNA, and polysaccharides such as pectin and xyloglucan from aqueous solutions by adding salt and ethanol as an antisolvent. In DNA extraction, after separating DNA from other cell constituents in water, DNA is precipitated out of solution by neutralizing it with positively ...