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  1. Antibody Guide

    Type
    Guide
    ... are called recombinant antibodies. Recombinant antibodies are created by cloning antibody genes from ...Science Guides Antibody Guide Antibody Guide View Addgene's Antibody Collection Jump To: Antibody Guide Production... produce antibodies, with the highest variability in antibodies produced. Cells - Antibodies can also ...amounts of a single antibody with little variation. Recombinant plasmids - Antibodies can be produced from...variable region and constant regions of the antibody to be independently manipulated. Antibodies produced this... in a buffer. Antibody Structure Figure 1: Structure of an antibody A standard antibody is made up of ...producing antibodies, can change the constant region, and therefore change the isotype of antibodies they ...
  2. Molecular Biology Reference

    Type
    Guide
    ...would not survive antibiotic treatment. It is important to distinguish that the antibiotic resistance gene...which is an important characteristic of why plasmids are convenient and easy to use. Antibiotic Resistance... grow in or on media containing that antibiotic. These antibiotic resistance genes not only give the scientist...information relating to antibiotic resistance genes as well as additional antibiotics not listed in the table...to confirm which antibiotic(s) to add to your LB media or LB agar plates. Antibiotic Recommended Stock...of ampicillin. Preparing Antibiotics Create a stock solution of your antibiotic. Unless otherwise indicated...Plasmids E. coli strains for propagating plasmids Antibiotics commonly used for plasmid selection DNA sequencing...
  3. CRISPR Guide

    Type
    Guide
    ...binding) SpCas9 VRER variant 3' NGCG SpCas9 EQR variant 3' NGAG SpCas9 VQR variant 3' NGAN or NGNG xCas9...LbCpf1) 5' TTTV AsCpf1 RR variant 5' TYCV LbCpf1 RR variant 5' TYCV AsCpf1 RVR variant 5' TATV Campylobacter...PAM sequence often found in plants, expanding the targeting range for plant genome engineering. Read more...more about Cas9 variants . Anti-CRISPR Prolonged Cas9 activity can increase the risk of off-target cutting...like 3xFLAG-tag, PA, and biotin tags, or an anti-Cas9 antibody. The CAPTURE system, developed by the Jian...Species/Variant of Cas9 PAM Sequence* Streptococcus pyogenes (SP); SpCas9 3' NGG SpCas9 D1135E variant 3' ...Journal . 2 (6), 376–394. PMID: 31742433 Durrant, M. G., Fanton, A., Tycko, J., Hinks, M., Chandrasekaran...
  4. Optogenetics Guide

    Type
    Guide
    ... its variants allow for inhibition of neurons using blue-green light. Mac Variants. Mac variants have ...wild-type opsin of each and some examples of relevant variants. This list is not exhaustive - please browse...Inhibitory (Hyperpolarizing) ChR Variants. Alternatively, ChR variants that inhibit neurons have been created... these variants result in the hyperpolarization of neurons. Examples of anion channel variants from other...thus inhibiting function of the neuron. NpHR Variants. Variants have been engineered with enhancements such... triggered by green-yellow light. Arch Variants. Arch variants have been developed with the following ...plasmids . Glossary of Microbial Opsin Variants Opsin Type Variant Description Peak Response Spectra (nm...
  5. Cloning

    Type
    Guide
    ... is important to confirm that there are no additional sites present within the fragments you want to assemble...Stitching Molecular cloning or the creation of recombinant DNA is an essential process used in scientific... most popular cloning methods used to create recombinant DNA. Restriction Enzyme Cloning Restriction enzyme... and ligated into your desired plasmid. It is important to note that restriction enzyme target sites can... fresh PCR product into a plasmid. The major disadvantage of TOPO® cloning is that very few plasmid backbones... known as Gibson assembly ( protocol ), takes advantage of the properties of 3 common molecular biology...chosen orientation, and without the need for any unwanted sequence at the junctions (such as a restriction...
  6. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...The mutant AAV-DJ8 displays the properties of AAV-DJ, but with enhanced brain uptake. AAV Variants As ...Aug;11(4):442-447. PubMed . AAV variants: Self-complementary recombinant adeno-associated virus (scAAV)...Uses Viral Integration Serotypes Pseudotyping AAV Variants References AAV Components The small (4.8 kb) ssDNA...lentiviruses integrate into the genome. This is important because local chromatin structure at the site ...is necessary for lytic cycle activation. When recombinant AAV (rAAV) is used for research purposes, the...expression in non-dividing cells and is a key advantage of rAAV. AAV Serotypes Eleven serotypes of AAV...hybrid viruses with new properties. Other system variants include: Self-complementary AAV (scAAV): One downside...
  7. Plan Your Experiment

    Type
    Guide
    ...knockout )? Generate a specific mutant allele of a gene ( point mutant )? Increase or decrease expression...desired sequence, you may want to consider alternative Cas enzymes ( see Cas9 variants and PAM sequences ).... specific disease or process of interest. Do you want to: Generate complete and permanent loss of gene...Cas9-gRNA ribonucleoprotein (RNP) complexes are advantageous due to the transient Cas9 expression. The table...may alter protein function when they occur in important protein domains. For gene editing experiments ... addition to off-target activity , it is also important to consider factors that maximize cleavage of ...assay (for NHEJ repaired DSBs): Provides a semi-quantitative readout of the percentage of alleles that have...
  8. Chemogenetics Guide

    Type
    Guide
    ...ligands can have high potency, which is potentially important for dosing in animal studies, made them attractive... N ‐oxide (CNO). CNO is a metabolite of the antipsychotic clozapine and seems to be pharmacologically ...however, it has off-target effects. The atypical antipsychotic drug olanzapine has also been shown to be an...domain was engineered to respond specifically to the anti-smoking drug varenicline. This PSAM carries mutations...consider: Chemogenetic activation or inhibition. Do you want to turn ON or turn OFF neurons in your experiment...potentially express the chemogenetic receptor in unwanted cell types. Generating a FLEx switch to control...until a cell or tissue-specific Cre is provided. Want to know more about FLEx vectors? Read our Plasmids...
  9. Lentiviral Guide

    Type
    Guide
    ...gene, conferring antibiotic resistance to infected host cells. When these antibiotics are added to the...and systems. When planning your experiment, the important component to consider and optimize is the transfer... media change and a brief incubation period, supernatant containing the virus is removed and stored or...order to create functional lentiviral particles. IMPORTANT : A 3rd generation transfer plasmid can be used...selectable markers conferring resistance to an antibiotic, but do encode another marker, such as GFP. A...integration site preferences. A major cellular determinant is LEDGF/p75, a lentiviral tethering protein ...with those in the vector into which one ultimately wants to ligate the insert. Often times it is less time...
  10. Science Guides

    Type
    Guide
    ... below. Antibodies Chemogenetics CRISPR Optogenetics Pooled Libraries Viral Vectors Antibodies Produced...system, antibodies are proteins capable of highly specific binding to other proteins. Antibody-based applications... often use antibodies conjugated to a signaling molecule, can be used to capture, quantify, and identify...
  11. Adenovirus Guide

    Type
    Guide
    ...in a higher background of non-recombinant adenoviral plasmids. Recombinant adenoviral plasmids are then...Coxsackie-Adenovirus Receptor (CAR) to enter cells. Recombinant adenovirus has two genes deleted: E1 and E3. ...recombination have not occurred. Once verified, the recombinant plasmid is linearized with PacI to create a linear...single-stranded DNA parvovirus and was discovered as a contaminant of adenovirus preparations, whereas adenoviruses...has published their method for generation of recombinant adenoviruses: A Practical Guide for using the...
  12. Addgene's eBook Collection

    Type
    Guide
    ... Overlay Select Antibodies 101 New! October 2024 | First Edition About Antibodies 101 October 2024...contains practical introductions and helpful tips for antibody-based applications, from flow to IHC! Close About...
  13. Guide to Using Pooled Libraries

    Type
    Guide
    ...can help you begin to understand what genes are important to a certain phenotype, and allow you to design...Both types of screen employ a selection method relevant to the phenotype being studied. Examples of published...lentiviral CRISPR libraries include: resistance to the anti-cancer drug vemurafenib , resistance to Clostridium...
  14. Retrovirus Guide

    Type
    Guide
    ... virus is produced in just a few days. Another variant, Phoenix-gp, contains only Gag-Pol and enables ...γ-retroviruses differ from lentiviruses? There are a few important differences between engineered γ-retroviruses ...names? The nomenclature of the plasmid backbone is meant to be descriptive of the components found within...
  15. Promoters

    Type
    Guide
    ...corresponds to the mRNA transcript produced. The antisense strand is referred to as the template strand or...itself which helps to explain how many physically distant DNA sequences can affect transcription of a given...
  16. Educational Resources

    Type
    Guide
    ...your molecular biology research. Topics include: Antibiotic Concentrations for Bacterial Selection What is...
Showing: 1 - 16 of 16 results