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We narrowed to 1,020 results for: NTS;

Showing: 161 - 180 of 1020 results

  1. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    ...various mechanisms. The RNA guide is typically 20 nts and the genomic locus must often be proximal to a...off-target events There are several strategies to lower the predicted frequency of off-target events, many ... to ensure off-target events are below a given threshold for certain experiments where they can’t be tolerated...single cell clones to use for multiple downstream experiments. You only successfully generate 1 null clone ...single clone is low, but since all downstream experiments will rely on that clone not having any confounding...rate of 5% is far too high when considering human patients and the diseases associated with an elevated mutation...the way of ‘good enough’. Predicting off-target events As mentioned in the above section, there are gRNA...

  2. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    ...flanking primer binding sites (designed to have 20-30 nt length, ~60 oC annealing temperature, and ~50% GC...are influenced by temporally ordered biochemical events. How do RSMs work? The state of an RSM is defined...interrogate state with qPCR. The re-arrangement of DNA segments in each state of a register enables the design... of a set of chemicals. Specific examples of arrangements that should be able to record all permuted substrings...gene regulation programs. The software has two components - a database of pre-compiled register designs...

  3. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    ...generated from a long hpRNA trigger, only a single ~21-nt interfering RNA is produced using shRNA/amiRNA. Although...possible in plants outside of limited, single cell protoplast analysis. Instead, microscopic plants must be...consuming, as RNAi mutants with a reduced growth phenotype will require additional plants to have enough ...LOF studies, the high gene copy number found in plants makes traditional knockouts unattractive from a... In either pathway, short double-stranded RNA fragments (siRNAs) are produced from long dsRNA via an RNase...interpretation of RNAi. Fortunately, researchers studying plants and other organisms lacking this innate immunity...transcriptionally or post-transcriptionally. In plants, different dicer-like enzymes (DCL) process the...

  4. Optogenetics Guide

    Type
    Guide
    ...function of the neuron. NpHR Variants. Variants have been engineered with enhancements such as: Human codon optimization... light. Arch Variants. Arch variants have been developed with the following enhancements: Increased light... its variants allow for inhibition of neurons using blue-green light. Mac Variants. Mac variants have ...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...Guide to Channelrhodopsin Variants: Features, Limitations and Future Developments. Exp Physiol. 96(1): 19...wild-type opsin of each and some examples of relevant variants. This list is not exhaustive - please browse our...

  5. Lentiviral Vector Guide

    Type
    Guide
    ...laboratory, the non-required components have been removed, and the required components have been separated into... of all lentiviral plasmid components, see the Lentiviral Plasmid Elements table . In order to produce... gone through improvements over the years for efficiency and safety. These improvements have been divided...our lentiviral guide to learn about lentiviral components, generations, lentiviral production, and common...Pseudotyping Common Uses Safety Resources Plasmid Elements Glossary Lentiviruses are a type of retrovirus...stable delivery of various genes, mutations, or treatments into cells, and has been widely used in the research...require different isoforms of these packaging components. Therefore, lentivirus may not be efficiently...

  6. Antibody Guide

    Type
    Guide
    ...antibody fragments consisting of two scFvs bound to a CH3 domain. Fab fragments - Fab fragments are created... called the antigen binding fragments (Fab). The tips of the Fab fragments contain highly variable regions...Affinity Reagents There are several alternatives to antibodies, ranging from antibody fragments (shown ...Structure Isotypes Antigen Specificity Other Affinity Reagents Handling Visualization Signal Amplification Application...Multiplex Assays Selection Validation Affinity reagents are molecular tools used in the lab to specifically...foreign bodies, are the most commonly used affinity reagents. Antibodies are Y-shaped proteins, naturally produced...interest is introduced to the animal, which then mounts an immune response to it. Once the immune response...

  7. CRISPR Guide

    Type
    Guide
    ...multiple Cas9 variants, including high fidelity Cas9s, Cas9s with various PAM requirements, and other Cas...scale edits by integrating segments of DNA flanked by specific repeat elements into the genome. CRISPR-associated...approach. Engineered CRISPR systems contain two components: a guide RNA ( gRNA or sgRNA ) and a CRISPR-associated...mechanisms of CRISPR and some of the ways the CRISPR components are leveraged to achieve specific goals, such...the Cas enzyme. Basic Mechanisms of CRISPR The requirements for a CRISPR knockout assay are simple. You’...mechanism. Multiplex Genome Engineering Many CRISPR experiments involve editing more than one gene. In these ...RuvC and HNH, can be altered to reduce cleavage events to a single cut (nick) or no cuts. Cas9 nickase...

  8. Molecular Biology Reference

    Type
    Guide
    ...promoters, small RNAs, and other genetic elements. Plasmid Elements Plasmids come in many sizes and vary ...amplified. These DNA fragments are attached to a glass slide so that different fragments of DNA, or templates... Recombinant DNA Working with Plasmids Plasmid Elements Types of Plasmids Molecular Cloning Common E. ...enable the isolation and purification of cellular components, such as DNA, but in practice this isolation ...restriction enzymes would yield about two million DNA fragments, which is far too many to isolate one specific...enabling the preparation of more manageable DNA fragments. In 1952, Joshua Lederberg coined the term plasmid...extrachromosomal heritable determinant. Plasmids are fragments of double-stranded DNA that typically carry genetic...

  9. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...gamma-retroviral plasmid components, see the Gamma-Retroviral Plasmid Elements table . In order to produce...gamma-retroviral guide to learn about gamma-retroviral components, retroviral production, and common uses. Science...Pseudotyping Common Uses Safety Resources Plasmid Elements Glossary Gamma-retroviruses are a subtype of retrovirus...stable delivery of various genes, mutations, or treatments into cells, and has been widely used in the research...require different isoforms of these packaging components. Therefore, gamma-retroviruses may not be efficiently...production, and scientists have made changes to these components for increased safety when producing viral vectors...produce gamma-retroviral vectors, these required components have been separated into distinct plasmids for...

  10. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...For a summary of all AAV plasmid components, see the AAV Plasmid Elements table . Figure 2: Plasmids required...random integration events can occur at a very low frequency. As with most experiments, infection risks ... viral vector guide to learn about AAV vector components, production and common uses. Science...AAV Types Common Uses Safety Resources Plasmid Elements Glossary Adeno-Associated Viruses (AAV) are small... necessary cellular machinery to produce large amounts of rAAV vectors. The E1 gene is not included in... Rep68, which can bind to specific Rep-binding elements present in the AAV ITRs and the AAVS1 site. Since...vectors. AAV Types Serotypes AAV serotypes are AAV variants that differ in their tropism — that is, the types...

  11. Adenovirus Guide

    Type
    Guide
    ... of all adenoviral plasmid components, see the Adenoviral Plasmid Elements table . Adenoviral Generations...random integration events can occur at a very low frequency. As with most experiments, infection risks ...titers for their use in experiments. Needle sticks and ripped gloves are common points of entry. Biosafety...vector guide to learn about adenoviral vector components, production and common uses. Science...Adenoviral Types Common Uses Safety Resources Plasmid Elements Glossary Adenoviruses (AdV) are medium-sized, ... cold. The immune system recognizes adenoviral elements rapidly and can mount a strong immune response...cells throughout viral vector production. During experiments, GFP can be used to sort cells infected with ...

  12. Plan Your Experiment

    Type
    Guide
    ...proteins available, and researchers can adapt these experiments for different organisms. This page provides a...CRISPR in your research and setting up CRISPR experiments. We will focus on making single edits using CRISPR...using CRISPR in other organisms. As with most experiments, you first need to have a biological question...you are ready to start designing the specific components (Cas enzyme and guide RNA) for your experiment...genetic manipulations require different CRISPR components. Selecting the genetic manipulation you plan ...plan to perform will help you narrow down which components you’ll need for a given experiment. In the table... window from the PAM sequence, and base editor variants often have different optimal editing windows. ...

  13. Molecular Cloning Techniques

    Type
    Guide
    ...40 bp of homology to each of the larger fragments. The fragments should have complementary ends with 30...restriction enzymes to cut dsDNA of interest into fragments containing precise 5' or 3' single-strand overhangs...and produce predictable cleaved ends in the DNA fragments. Given its prevalence, the vast majority of plasmids...polymerase, and ligase. In Gibson assembly, DNA fragments with 20–40 base-pair homology at their ends can...exonuclease chews back the 5’ ends of your DNA fragments, generating long overhangs that anneal to each...is that it allows for the assembly of multiple fragments of DNA in the chosen orientation at a time, and...enzyme or Gateway recombination sites). Any dsDNA fragments can be used, so if properly designed, any insert...

  14. Promoters

    Type
    Guide
    ...with BioRender.com. DNA sequences called response elements are located within promoter regions, and they ...regulatory sequences, including enhancers, boundary elements, insulators, and silencers. Bacterial Promoters.... It is not unusual to have several regulatory elements, such as enhancers, several kilobases away from...nucleosomes) also bind the TATA box. Histone binding prevents the initiation of transcription, whereas transcription...promoter, which contains many primary regulatory elements. The proximal promoter is found approximately ... binding sites, but mostly contains regulatory elements. Common Eukaryotic Promoters Researchers often...

  15. Guide to Using Pooled Libraries

    Type
    Guide
    ...you to design additional hypothesis-directed experiments. As you might have guessed from their name, pooled... false positives or false negatives in later experiments, and can also negatively affect data reproducibility...only way to accurately evaluate which library components are influenced by selection, so it’s essential...allows for parallel sequencing of millions of DNA fragments simultaneously in a single reaction. As discussed...

  16. Chemogenetics Guide

    Type
    Guide
    ...barrier in rats, and the utility of DREADDs in experiments in this species may be entirely due to back-metabolism...issues as CNO and are attractive alternatives for experiments. Perlapine has been previously used in human ...respond to the same ligands was that bidirectional experiments (i.e. using both activating and inhibiting DREADDs...absence of Cre and are especially useful in AAV experiments where the virus can spread around the injection...

  17. Addgene's eBook Collection

    Type
    Guide
    ...applications, tips for using vectors in your experiments, troubleshooting advice on production and use...

  18. Gibson Assembly Protocol

    Type
    Protocol
    ...Join almost any two fragments regardless of sequence. No scar between joined fragments. Fewer steps. One...about what DNA segments you will need to join to create your final plasmid. Adjacent segments should have...If there are significant amounts of undesired product, gel-purify DNA segments. Otherwise, PCR purification...combine segments in the Gibson Assembly Reaction. Pro-Tip Yields will be best when the DNA fragments are ...annealed DNA fragments. Taq DNA Ligase - covalently joins the annealed complementary DNA fragments, removing... al. , 2010). Number of fragments assembled simultaneously Multiple fragments can be assembled in one ... for the easy assembly of multiple linear DNA fragments (Gibson et al. , 2009). Regardless of fragment...

  20. Pipetting Protocol

    Type
    Protocol
    ...bottom. For the P1000, this represents the ones. For the P200, this represents the first decimal place. ...For the P20, this represents the second decimal place, and for the P2 this represents the third decimal...in many scenarios to accurately dispense small amounts of liquid (think: 0.1 µL to 1 mL). When working...microfuge tube, bottle, etc.) Labels for containers Reagents Liquid for pipetting Background Information Anatomy... container in order to avoid contaminating the contents. Once the end of the tip is submerged, slowly ...small changes in the volume could affect your experiments. Beyond knowing the proper way to pipette, it...they are properly tuned to dispense the correct amounts of liquid. If you are pipetting the same amount...
Showing: 161 - 180 of 1020 results