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We narrowed to 810 results for: mal

Showing: 761 - 780 of 810 results

  1. 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...

  2. 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...

  3. 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...

  4. 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...

  5. 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 ...

  6. 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...

  7. 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...

  8. 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...

  9. 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...

  10. 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...

  11. 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...

  12. 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...

  13. 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...

  14. 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...

  15. Protocol - Over-Agar Antibiotic Plating

    Type
    Protocol
    ...through the use of a selection curve to determine optimal antibiotic concentration. Last Update: Oct. 27,...pipetting and spreading Bunsen burner (or other small flame source) Incubator Reagents 6 cm diameter LB... the liquid is mostly absorbed (there is a very small visible volume of pooled liquid remaining on the...over-agar Plate shows several individual colonies with smaller size than the 1 mg/mL and 2 mg/mL plates and effective...

  16. Protocol - Bacterial Transformation

    Type
    Protocol
    ... nearly all plasmids (even those designed for mammalian cell expression) carry both a bacterial origin... cells, which come frozen and are prepared for optimal transformation efficiencies upon thawing. For the...important if you will be transforming with very small amounts of DNA or if you're multiple plasmids at...the cells by centrifugation and resuspend in a smaller volume of LB so that there isn't too much liquid...

  17. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...for 1 minute at 72°C: The Taq polymerase has an optimal temperature around 70-75°C so this step enables...creating a “master mix,” which minimizes the number of small volumes you need to pipet. If you are using the ...needs to reconstitute their primers in liquid, normally sterile dH 2 O. To make a 100uM stock of any primer...efficiently. Taq Buffer with MgCl 2 : Provides an optimal and stable chemical environment for the DNA polymerase...

  18. Science Guides

    Type
    Guide
    ...pathways. Receptors are remotely controlled via small molecules, which allows for specific control of ...

  19. Lentivirus ddPCR Titration

    Type
    Protocol
    ...kDa subunit (RPP30) are used as a control for normalization. The dilution series outlined in this protocol...5425R Droplet digital PCR System, Bio-Rad, DX200 Thermal Cycler, Bio-Rad, T100 PCR Plate Sealer, Bio-Rad...plate has been sealed, proceed to thermocycling. Thermal Cycling Run the following PCR parameters: Cycling...risk of contaminating reagents we recommend making small aliquots of master mixes, and primers/probe mixes...

  20. Video Library

    Type
    Protocol
    ...Deletions with CRISPR Generate genomic deletions in mammalian cells lines using CRISPR/Cas9 Genomic Deletions...adeno-associated virus from a preparation produced in mammalian cells AAV Purification by Iodixanol Gradient Ultracentrifugation...culture in the lab. Blog post: 10 Basic tips for mammalian cell culture Aseptic Technique Aseptic technique...
Showing: 761 - 780 of 810 results