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Showing: 121 - 140 of 567 results

  2. Zhang Lab CRISPR Page

    Type
    Collection
    ...20bp). Return to top References SpCas9 plasmids Multiplex Genome Engineering using CRISPR/Cas Systems. Cong...

  4. Genetic Code Expansion

    Type
    Collection
    ...typically expressed from a single plasmid, with multiple copies of the tRNA. Applications By making small...

  5. Trimmer Lab NeuroMab Collection

    Type
    Collection
    ...subclass from the conventional mAb to facilitate multiplex labeling with subclass-specific secondary antibodies...recombinant monoclonal antibodies for enhanced multiplex immunolabeling of brain. eLife . 8:e43322. PMID...

  6. Molecular Biology Reference

    Type
    Guide
    ...Sequencing (NGS) comes in. NGS is a high-throughput, multi parallel sequencing platform that can generate sequencing...combinations each amino acid is encoded for by multiple codons. Plasmids and Recombinant DNA Technology... as cloned DNA, and this process of generating multiple identical copies of a recombinant DNA molecule...the bacterial host. Each bacterium can contain multiple copies of an individual plasmid, and ideally would...bacteria by bacterial transcriptional machinery. Multiple Cloning Site (MCS) Short segment of DNA which ...pressure to maintain and replicate your plasmid over multiple generations. More information relating to antibiotic...puzzle pieces in only a few reactions.There are multiple approaches to acquire NGS but one of the most ...

  7. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ... single channel pipette 20–200 µL multichannel pipette Multichannel pipette reagent reservoirs Pipette... and set aside. Aspirate the wells and use a multichannel pipette to add 200 µL wash buffer to each well...tube and invert several times to mix. Using a multichannel pipette, add 200 µL of blocking buffer to each...invert the tubes several times to mix. Using a multichannel pipette, add 100 µL of primary antibody solution...invert the tube several times to mix. Using a multichannel pipette, add 100 µL of secondary antibody to...

  8. AAV ddPCR Titration

    Type
    Protocol
    ...pipette 1–10 µL multichannel pipette 2–50 µL multichannel pipette 20–200 µL multichannel pipette Vortex...Use multichannel pipettes for the dilution series. For dilutions 1–5, use the 1–10 µL multichannel pipette...polystyrene reagent reservoir. Using a 20–200 µL multichannel pipette, carefully add the dilution buffer to...to 5 µL. For dilutions 6–8, use the 20–200 µL multichannel pipette set to 50 µL. For mixing between dilutions...dilutions, use the 20–200 µL multichannel pipette set to 90 µL and mix by pipetting the liquid up and down...cartridge into the cartridge holder. Using a 2–50 µL multichannel pipet, load 20 µL of the reaction mixtures into...polystyrene reagent reservoir. Using the 20–200 µL multichannel pipet, load 70 µL of droplet generation oil ...

  9. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ...pipette 1–10 µL multichannel pipette 2–50 µL multichannel pipette 20–200 µL multichannel pipette Reagents...i.e master mix + water Whenever possible use a multichannel pipet to minimize pipetting error and variability... samples very well by pipetting back and forth multiple times at each step Reagent Preparation Master ...immediately before use. Use a reservoir and a multichannel pipette to dispense the master mix into the ...time to make a new one. When developing the assay multiple plasmids containing ITR were tested. Plasmid #59462... results. Use the recommended plasmid, or test multiple plasmids to find a suitable one. Some labs have...with >50 uL if your well contains 100 uL) Use a multichannel pipette to load the standards and samples onto...

  10. Polymerase Chain Reaction Overview and Applications

    Type
    Blog Post
    ...Quantification.”   Multiplex PCR Multiplex PCR, as the name implies, is a method in which multiple targets can...forward and reverse primer sets.  Multiple template PCR - multiple templates with different primer pairs...Scientists can also utilize multiplex qPCR to quantify the concentrations of multiple starting DNA templates... changes in target DNA levels Multiplex PCR Amplify multiple targets from one sample Disease or...practice in molecular biology and can be used in a multitude of scientific techniques such as molecular cloning... be amplified in a single PCR experiment using multiple primers all in one PCR reaction. This is an extremely...laboratory.  There are two main categories of multiplex PCR:  Single template PCR reaction - one template...

  11. Deep Dive: qPCR

    Type
    Blog Post
    ... If there are multiple amplicons, or primer dimers forming, the curve will show multiple peaks.    ...optimizing for LightCycler machines.) Multiplexing It is possible to multiplex qPCR when using fluorescent probe-based...change, the multiplexed protocol may have to be re-optimized.  While troublesome, multiplexing your reference...the Lightcycler while being relatively easy to multiplex (Thellwell, 2000). See Table 1 for a comparison...noise Probe Allelic specificity Can be multiplexed Single amplicon validation technique Hydrolysis... reagents in your chemistry, unless running a multiplex reaction or on a rapid cycle machine. Melt Curves...data will be much improved if you move to using multiple reference genes whenever possible (Vandesompele...

  12. Rewiring Metabolic Circuitry with CRISPR RNA Scaffolds [Video]

    Type
    Blog Post
    ...transcriptional programs on multiple genes. Modifying the CRISPRi system for multidirectional control The CRISPR...coli. The utility of simultaneous and multidirectional multiplex control via scRNA design is readily apparent...targeted site. Multiple scaffold RNAs can be designed to allow simultaneous, multidirectional and tunable...discussed in the paper. The transcriptional control of multiple loci is deftly coordinated by the eukaryotic cell...toward tinkering with regulatory networks and multiplex gene control. However, these approaches are often...Modifying the CRISPRi system for more flexible multidirectional control, a new study repurposes the sgRNA ...scaffold modules are comparatively small allowing multiple sgRNA constructs to be introduced simultaneously...

  13. Lentivirus ddPCR Titration

    Type
    Protocol
    ...single channel pipette 2–50 µL multichannel pipette 20–200 µL multichannel pipette Ice bucket 96-well freezer...cartridge into the cartridge holder. Using a 2–50 µL multichannel pipet, load 20 µL of the reaction mixtures into...polystyrene reagent reservoir. Using the 20–200 µL multichannel pipet, load 70 µL of droplet generation oil ...droplets have been generated, use a 20–200 µL multichannel pipette to collect 40 µL of droplets. Pro-Tip... RPP30 in diploid cells, thus the reason for multiplying by 2. $$V = 2*{copies\ RRE \over copies\ RPP30...

  14. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    ...visualize multiple targets at once. An advantage of multiplexing is that you can ask how multiple proteins...and colors makes it easy to perform multiplexed IF. By using multiple primary antibodies made in different...antibodies are generally polyclonal, which means that multiple secondary molecules can bind to a single primary...species and to label targets with different colors. Multiplex immunofluorescence Speaking of different colors... excitation/emission spectra do not overlap. Multiplexed IF can also require some additional optimization...antibodies. But, once you do find the right protocol, multiplex IF can give you impactful and beautiful results...minimal impact to the target epitope. There are multiple methods for fixing samples, each with its own ...

  15. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    ...for Biological Studies) Multiple uses: From multiplexing to multiscaling NIR-shifted spectrum is perfect...perfect for use in spectral multiplexing (Shcherbakova et al., 2018). NIR FPs can be combined with blue-light...These probes can be combined in two-color and multicolor NIR imaging.  Small miRFPnano A separate class...Verkhusha, V.V. Near-Infrared Fluorescent Proteins: Multiplexing and Optogenetics across Scales. Trends Biotechnol...fluorescent proteins as tags and biosensors for multiscale imaging. Nat Commun 7, 12405 (2016). Qian, Y....monomeric near-infrared fluorescent proteins for multicolor imaging across scales. Nat Commun 11, 239 (2020...cyanobacteriochrome as versatile tag for spectral multiplexing. Nat Commun 10, 279 (2019). Oliinyk, O.S. et...

  16. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ...Combining multiple modifications A key feature of IVA cloning is the ability to combine multiple modifications...vivo to produce the multiply modified product. For example, by combining multiple primers, a C-terminal...GFP-tag (Figure 3). Achieving multiple modifications requires multiple recombination events to assemble...regions can be encoded in the primer sequences. Multiple plasmids can be included in a single PCR, for ...of the strain: when asking the cell to perform multiple recombination events, all linear fragments need... 107 CFU/μg are perfectly fine, yet performing multiple modifications will require ultracompetent (109..., and point mutagenesis to complex assembly of multiple genes. Primer design for each type of modification...

  17. CRISPR-based Genome Editing Systems For Editing in E. coli

    Type
    Blog Post
    ...for multicopy chromosomal integration (ex: for protein expression) (Zhang et al., 2020). Multicopy chromosomal... Most recently, we’ve developed a new system “Multicopy chromosomal integration using CRISPR-associated...including concatenations into crRNA arrays targeting multiple different sites in E. coli. pCutamp cures the ...Ptet, and Para. The pDonor plasmid pRE57-Ter with multiple terminators can serve to interrupt genes (or gene.... Figure 2: (A) MUCICAT plasmids for multicopy integration. (B) Anhydrotetracycline (aTc) induces...following advantages: Fast and site-specific multicopy chromosomal integration of cargo without using...dosages to facilitate swift optimization of strains. Multiple induction systems to choose from. Ready for metabolic...

  18. Antibody Guide

    Type
    Guide
    ...five human isotypes. Some isotypes naturally form multi-antibody units, while others exist as monomers. ...antibody in assays with multiple targets, and is described in more detail in the Multiplex Assays section below...differentiation between multiple antibodies. They are often used in assays that have multiple proteins to detect... controls. Multiplex Assays Flow cytometry, IF, IHC, and ICC are typically run as multiplex assays. Both...Methods Visualization Methods Cell Sorting Methods Multiplex Assays Selection Validation Affinity reagents ...allows for the amplification of signals by having multiple secondary antibodies binding to a primary antibody..., which is typically the number of antibodies multiplied by two, as each antibody has two binding sites...

  19. Antibodies 101: Secondary Antibodies

    Type
    Blog Post
    ... but it probably should be. In a multiplex assay - one with multiple proteins of interest - the secondary... simplifying multiplex assays.  A brief note Since the indirect method relies on multiple secondary antibodies...antibody instead of the primary antibody. Since multiple secondary antibodies can bind to a single primary...stem of an antibody’s Y) is large enough to allow multiple secondary antibodies to bind to a primary antibody...antibody protocols are designed to take place over multiple days. Increased run time is considered one of ...it except in the most dire of circumstances.) Multiplex assays Mo’ proteins, mo’ problems may not be a...different species and/or isotypes. For highly multiplexed assays, using secondary antibodies which recognize...

  20. Deep Dive: Statistical Tests (Comparisons)

    Type
    Blog Post
    ...on crop production.   MANOVA The MANOVA is a multivariate form of ANOVA. Isn’t that what a two-way ANOVA.... Factorial ANOVA This type of ANOVA assesses multiple independent variables on a single dependent variable...variables are independent of each other. That is, multiple independent variables are assumed to not have ...Knowing that significance exists somewhere amongst multiple groups is rarely enough information. A post hoc...performed a MANOVA/MANCOVA and want this information, multiple ANOVAs/ANCOVASs must be performed first, and then... by the total number of comparisons performed multiplied by the alpha value is less than or equal to the...combinations – Tukey’s test, Newman-Keuls, Duncan’s multiple range test, and several others. Each test has ...
Showing: 121 - 140 of 567 results