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TypeGuide...performs a chemiluminescent reaction in the presence of several different substrates. The luminescence produced...Recombinant antibodies are created by cloning antibody genes from specific hybridoma cells into plasmids. Plasmids...region is a stable part of the antibody, which determines the isotype of the antibody. B cells, the immune...tears, saliva, mucus) where it protects mucosal membranes Dimer with a valency of 4 IgA1; IgA2 IgD Coexpressed... epitope, or polyclonal, meaning there are many clones that are specific to multiple epitopes on one antigen...Sodium azide, an antimicrobial agent, can block some amines needed for conjugation reactions and should be ... tubulin (sometimes referred to as housekeeping genes). This allows for relative comparison of expression...
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Guide to Using Pooled Libraries
TypeGuide... multiple genes at once. A well-designed screen can help you begin to understand what genes are important...genetic screening, or identifying previously unknown genes that contribute to a phenotype. At their simplest... backbone, but they express or target different genes. Some libraries cover the majority of the genome... if they are designed to cover only a subset of genes, or very large (e.g. the Toronto KnockOut (TKO) ...Sequence those cells that pass (“win”) Get a list of genes involved in the selection NEGATIVE screen: Apply...generally very robust, and tens of thousands of genes that were initially present may be narrowed down...screen, negative screens are often used to identify genes that are essential for growth/survival under certain... -
Promoters
TypeGuide...general transcription factor proteins and histones can bind. Histones are proteins found in eukaryotic cells...transcription of genes. There are three types of RNA polymerases that all transcribe different genes. RNA polymerase...like operons. Operons are a cluster of different genes that are controlled by a single promoter and operator...repressor or activator can bind, and the structural genes that are transcribed together. Operon regulation...polymerase I transcribes genes encoding ribosomal RNA (rRNA) which is a main component of a cell’s ribosome...translation of a protein. RNA polymerase III transcribes genes encoding transfer RNAs (tRNA), the adaptor molecules... -
Plan Your Experiment
TypeGuide...more effective than dCas9 alone for mammalian cell lines Interfere Activate (CRISPRa) Increase gene expression...can be transient, or you can generate stable cell lines if your plasmid contains a selection marker. Plasmid..., note that this method is most useful for cell lines that you can transfect at high efficiency, such ... are a popular option for producing stable cell lines, since the genetic cargo (in this case, CRISPR components...is a common choice for generating modified mouse lines. Protein delivery Lastly, you can deliver Cas-gRNA...Elongation roadblocks mediated by dCas9 across human genes modulate transcription and nascent RNA processing... -
Chemogenetics Guide
TypeGuide... PSAM plasmids ! LMOs Luminopsins (LMOs), or luminescent opsins, are opto-chemogenetic fusion proteins... LMOs a versatile option. Coupling brighter bioluminescent modules, such as luciferase-fluorescent protein... a single molecule driven by FRET-modified bioluminescence. Neurophotonics. 11:021005. PMID: 38450294 ...Manipulation for Ameliorating Huntington's Disease Pathogenesis. iScience , in press. Magnus CJ, Lee PH, Atasoy...Inhibitory luminopsins: genetically-encoded bioluminescent opsins for versatile, scalable, and hardware-independent... -
Optogenetics Guide
TypeGuide... chimera 470 ChIEF, ChEF, ChD Engineered by chimeragenesis of ChR1 and ChR2 450-470 C1V1 ChR1-VChR1 chimera...injection site. Optical Switches Researchers have harnessed plant and bacterial photoreceptors to create protein...improve upon previous tools through directed mutagenesis - use Addgene's Optical Switch Plasmid Table ... sequester and release proteins from cellular membranes using light. light-induced protein dissociation... -
Cloning
TypeGuide...molecular cloning scientists can amplify and manipulate genes of interest and then insert them into plasmids for...putting it under a variety of promoters and into backbones with different selection cassettes). Although ...disadvantage of TOPO® cloning is that very few plasmid backbones are available TOPO® ready, and it is not feasible... -
Educational Resources
TypeGuide...CRISPR Viral Vectors Plasmid Technology Career Readiness Molecular Biology Learn about plasmids, sequencing... -
Science Guides
TypeGuide... backbone, but they express or target different genes. Some libraries cover the majority of the genome...