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We narrowed to 816 results for: MAL

Showing: 761 - 780 of 816 results

  1. Antibody Guide

    Type
    Guide
    ...methods: in animals, in cultured immune cells, or through the use of recombinant plasmids. Animals - Antibodies...Antibodies produced in animals make use of the animal’s immune system. The antigen of interest is introduced...introduced to the animal, which then mounts an immune response to it. Once the immune response has developed...antibody. Monocultures of B cells isolated from animals immunized as described above are used to create... protein chains: two large heavy chains and two small light chains. These four domains come together to...only produce one variable region. Isotypes The mammalian immune system produces five classes of antibodies...antibodies are typically collected directly from animals and contain antibodies targeting various epitopes...

  2. Promoters

    Type
    Guide
    ... TATAAT. Proximal Promoter Further upstream from the core promoter you will find the proximal promoter...Constitutive Strong mammalian promoter from human cytomegalovirus EF1a Constituitve Strong mammalian promoter from... Constitutive Strong hybrid mammalian promoter PGK Constitutive Mammalian promoter from phospholycerate...portions that make up a promoter: core promoter, proximal promoter, and distal promoter. Below describes...Promoter The core promoter region is located most proximal to the start codon and contains the RNA polymerase...contains many primary regulatory elements. The proximal promoter is found approximately 250 base pairs...called the distal promoter which is upstream of the proximal promoter. The distal promoter also contains transcription...

  3. Molecular Biology Reference

    Type
    Guide
    ...order to isolate a particular gene from human chromosomal DNA, it would be necessary to isolate a sequence...enables the preparation of more managable (i.e., smaller) DNA fragments. In 1952, Joshua Lederberg coined...coined the term plasmid, in reference to any extrachromosomal heritable determinant. Plasmids are fragments...carry genes and can replicate independently from chromosomal DNA. Although they can be found in archaea and...relationship with the host cell. Like the bacterial chromosomal DNA, plasmid DNA is replicated upon cell division...plasmids are replicated in E. coli and are relatively small (∼3000 - 6000 basepairs) to enable easy manipulation...fragments can be inserted. When a plasmid exists extrachromosomally in E. coli , it is replicated independently...

  4. Modular Cloning Guide

    Type
    Guide
    ... Assembly into acceptor vectors. The Mammalian Toolkit Mammalian Expression Hana El-Samad 253 plasmids...MoClo-YTK . EMMA: Extensible Mammalian Modular Assembly Toolkit Mammalian Expression Yizhi Cai 95 plasmids...generate mammalian expression vectors for transient or stable transfections. PUF Assembly Kit Mammalian Expression...homology domain, for mammalian or E. coli expression. T2A Split/Link TurboID Kit Mammalian Expression Sven ...constructs for integration into Y. lipolytica chromosomal loci. Yeast GPCR-sensor Toolkit Yeast Expression... Kit Bacterial Expression, Insect Expression, Mammalian Expression Stephan Gruber Platform for rapid assembly... Golden Gate-based cloning methods for use in mammalian cells. These can be used with plasmids from the...

  5. Plan Your Experiment

    Type
    Guide
    ...vector) is smaller than that of lentivirus. Thus, AAVs are generally only compatible with smaller Cas enzymes...focus on making single edits using CRISPR/Cas9 in mammalian cells as an example, but many of these principles...common categories of genetic manipulations used in mammalian cells and their corresponding Cas enzymes. In ...specific genomic edits, like point mutations or small tag insertions, by copying from a DNA donor template...exert their activity within a very narrow window proximal to the PAM sequence Base Edit Prime Edit Generate...KRAB can be more effective than dCas9 alone for mammalian cell lines Interfere Activate (CRISPRa) Increase... and base editor variants often have different optimal editing windows. Be sure to refer to the literature...

  6. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...Adeno-Associated Viruses (AAV) are small, non-enveloped viruses containing a small, linear single-stranded DNA...genome. Adeno-associated viruses (AAV), however, are small single-stranded DNA viruses that belong to the Parvoviridae...limited by the physical space available inside of the small AAV capsid and directly correlates to the size of...molecular weight structures that are maintained extrachromosomally. Random integration of rAAV genomes in non-homologous...period of years without damaging the host cell. Episomal stability allows for long-term transgene expression... deliver genes to a wide range of tissues with minimal immune response, and the low risk of insertional...neural connections and brain structure due to their small size, low immunogenicity, and limited ability to...

  7. Chemogenetics Guide

    Type
    Guide
    ...can be turned on or off by the application of a small molecule ligand. The ideal chemogenetic tools are...native ligands and are engineered to respond to small molecules that do not affect endogenous signaling...potency, which is potentially important for dosing in animal studies, made them attractive targets for development...muscarinic receptor. This DREADD was shown to have a small amount of constitutive activity, and is not widely...Compound 21. Both Compound 21 and DCZ seem to have minimal off-target activity, and do not seem to have the...receptor (nAChR) domains that respond to specific small molecules termed Pharmacologically Selective Effector...available. Additionally, genetically engineered animal models expressing hM3Dq, hM4Di, and rM3D DREADDs...

  8. Adenovirus Guide

    Type
    Guide
    ...Instead, they remain as extrachromosomal elements within the nucleus, normally as double-stranded circular...cause a range of mild diseases in humans and other animals, most commonly respiratory infections, such as ...genome. Adeno-associated viruses (AAV), however, are small single-stranded DNA viruses that belong to the Parvoviridae...needs to be expressed by the packaging cell line, normally the human embryonic kidney 293 (HEK293) cell line...transgene expression delivered through rAdV vectors is normally transient and short-lived. Second- and third-generation...excellent vectors for delivering target antigens to mammalian hosts and have been used as vaccines against infectious...severity is increased with rAdV vectors as they are normally produced in high titers for their use in experiments...

  9. Cloning

    Type
    Guide
    .... Back to Top Gibson Assembly (Isothermal Assembly Reaction) Isothermal cloning, more commonly known as...their ends can be easily ligated together in one isothermal reaction. First the 5’ exonuclease chews back... coli , where the nicks will be repaired by the normal replication process. It is important to note that...

  10. Guide to Using Pooled Libraries

    Type
    Guide
    ...plasmids in a single tube. Pooled libraries can be small, if they are designed to cover only a subset of ... their plasmids from the population, and only a small fraction survive. After the surviving cells are ... Company Resources Science Exchange Scientist (formally Assay Depot) Resources Find additional resources...

  11. Immunocytochemistry

    Type
    Protocol
    ... of cold 4% paraformaldehyde in PBS on ice for 15 min . Pro-Tip While 4% Paraformaldehyde fixation works...labeling HeLa cells for a target protein using the formaldehyde fixation method. The protocol may need to be...coated coverslips HeLa cells 24-well plate 4% Paraformaldehyde 5 mg/mL 4′,6-diamidino-2-phenylindole (DAPI...be better for some applications. Remove the paraformaldehyde and follow your institution's laboratory safety...concentration in antibody dilution buffer. Pro-Tip The optimal antibody concentration will vary but generally ...fluorescent filters. Tips and Troubleshooting The optimal fixation method will vary depending on the sample...

  12. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...Web resources Addgene’s mammalian RNAi website: www.addgene.org/mammalianrnai The RNAi Consortium (TRC...Designing shRNA Oligos for pLKO.1 B.1 Determine the optimal 21-mer targets in your gene B.2 Order oligos compatible... F.1 Recommended materials F.2 Determining the optimal puromycin concentration F.3 Protocol for lentiviral...resistance gene for selection of pLKO.1 plasmid in mammalian cells. sin 3’LTR 3’ Self-inactivating long terminal...’s lab. For more information, visit Addgene’s Mammalian RNAi Tools page. Several other laboratories have...Designing shRNA Oligos for pLKO.1 B.1 Determining the Optimal 21-mer Targets in your Gene Selection of suitable...Weinberg lab. For more information, visit Addgene’s Mammalian RNAi Tools page. E.2 Protocol for Producing Lentiviral...

  13. Pipetting Protocol

    Type
    Protocol
    ...represents the first decimal place. For the P20, this represents the second decimal place, and for the ...pipette in many scenarios to accurately dispense small amounts of liquid (think: 0.1 µL to 1 mL). When ...volume display on these pipettes will also have small tick marks at the bottom. For the P1000, this represents...the P2 this represents the third decimal place. Procedure How to Pipette Now that you know the different...pipetting from a relatively large container holding a small volume of liquid, tilt the container holding the...accurately and precisely is important since even small changes in the volume could affect your experiments...

  14. Optogenetics Guide

    Type
    Guide
    ... viral vector with a floxed opsin in a VGAT-cre animal will result in expression of the opsin only in ...optically assembled protein architectures in living mammalian cells. LITE Two-hybrid system integrating the ...

  15. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...envelope, Phoenix-AMPHO (for the infection of mammalian cells). Using this system, viral vectors are produced... G., Pond, S. M., Ferris, A. L., Hughes, S. H., Malech, H. L., & Wu, X. (2014). Enhancers are major targets...

  16. Western Blot

    Type
    Protocol
    ...buffer and optimal antibody concentrations. Consider titrating your antibody to determine the optimal dose.... concentration in blocking buffer. Pro-Tip The optimal concentration will vary between antibodies but ...detect the bands. Tips and Troubleshooting The optimal lysis buffer will vary depending on the sample ... buffers to find the best for your target. The optimal concentration and blocking buffer will vary between...consider titrating your antibody to determine the optimal dose. To ensure that your antibody is both functioning...

  17. Gibson Assembly Protocol

    Type
    Protocol
    ...overlapping DNA fragments can be joined in a single isothermal reaction. With the activities of three different...two-part Gibson reaction if you're only making a small change in a plasmid (such as point mutations). Generate...concentrations. The Gibson Assembly master mix (isothermal reaction mix) consists of three different enzymes...-Stranded DNA-Binding protein (ET SSB) to the isothermal reaction mix. ET SSB protects 3’ ssDNA ends from... in the Gibson reaction are different than with normal PCR primers (Gibson et al. , 2010). Number of fragments...Cepko C. (2020). A Simple Enhancement for Gibson Isothermal Assembly. bioRxiv . https://doi.org/10.1101/2020.06.14.150979...

  18. DNA Quantification

    Type
    Protocol
    ...your substance maximally absorbs. In the case of nucleic acid (DNA and RNA), the maximal absorbance is ...and most have a specific wavelength that they maximally absorb. By measuring the absorbance of a liquid...is at 260nm. Protein maximally absorbs at 280nm and the ratio of nucleic acid to protein (260/280) is generally...days, many labs have a NanoDrop, which is a very small spectrophotometer that can accurately read DNA concentration...

  19. Kit Free RNA Extraction

    Type
    Protocol
    ...While simple homogenization is effective for most mammalian tissues; more hardy tissues such as bone, or bacteria...Pro-Tips If you anticipate your RNA yield to be small, RNase-free Glycogen may be used as a carrier to...cycles of your entire RNA sample, consider making smaller aliquots of it and storing those in -80°C. Option...cycles of your entire RNA sample, consider making smaller aliquots of it and storing those in -80°C. Solution...

  20. CRISPR Library Amplification

    Type
    Protocol
    ...pressure. These recombined plasmids are much smaller and are a smaller nutritional burden on the bacteria. Bacteria...modifications dictated by the originating laboratory for optimal results. If you obtained the pooled library from...can recombine between their LTRs resulting in a smaller plasmid containing only the elements required for...convenience but any brand or house- made LB that supports normal growth is expected to work. 50 mL Falcon Conical...library of 100,000 plasmids. Colonies may appear small and require extra incubation time in order to be...
Showing: 761 - 780 of 816 results