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We narrowed to 1,078 results for: CAN

Showing: 381 - 400 of 1078 results

  1. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ... inexpensive and can be 5’-phosphorylated in house using a simple PNK reaction or can be purchased 5’-...required for its optimization and development before it can truly be pitted against CRISPR head-to-head.  Exciting...rather than an RNA guide as with Cas9. Third, it can be used to target “difficult” GC-rich regions of ...protein (probably while it is being synthesized) can form a complex with an ssDNA guide. Therefore, researchers...versatility of its delivery methods. Researchers can conveniently transfect/inject cells with in vitro...achieved by a few respondents suggesting that NgAgo can indeed be optimized for genome editing in mammalian...time is required for NgAgo optimization before we can make any major conclusions: That said, many people...

  2. Plasmids 101: Colony PCR

    Type
    Blog Post
    ... you can do this. If you are going to complete your colony PCR analysis in the same day, you can save ... on an LB plate. You can use this plate to start liquid cultures. Lastly, you can start small overnight...with restriction enzyme digest; however colony PCR can accomplish the same thing in less time and for less...clone without the insert. This type of primer pair can tell you if the insert is the correct size and whether...the size of the insert and if it's in the plasmid Can be used to screen clones that were created with the...Taq polymerase is sufficient. Controls: Controls can make or break an experiment. The best controls for... insert. These controls are quick references you can use when you run your PCR products out on a gel to...

  3. CRISPR Cheat Sheet

    Type
    Blog Post
    ...: Cut: CRISPR can be used to cut the DNA sequence. With a repair template, CRISPR can be used to introduce...100% specific and can cut DNA sequences that researchers don’t intend it to. CRISPR cannot cut all sequences...DNA. Fusing the nuclease to an adenosine deaminase can convert adenosine to inosine. Activate/Repress: Modified...proteins continue to expand the targeting range. CRISPR can be difficult to deliver. Not all cells efficiently... Modified versions of CRISPR nucleases that can’t cut DNA (dCas9, for example) that are used to increase...

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

    Type
    Blog Post
    ...function downstream of DNA, such as RNA interference, can reversibly exert their effect independent of gene...of different small RNAs (Figure 1). These siRNAs can exert their gene silencing effect through targeted...sequences (Figure 1). As a result, gene silencing can be easily induced by exogenous application of long...machinery. Alternatively, endogenous miRNA precursors can be engineered to created artificial miRNAs. The processed...CRISPR-generated mutants believed to be knockouts can maintain partial function due to the use of exon ...trigger RNAi activation. A typical RNAi experiment can be divided into three fundamental steps: creation... (Figure 1). With this technique, multiple genes can be simultaneously silenced if a consensus sequence...

  5. Mouse Modeling, Part 2: Breeding and Crossing Mice

    Type
    Blog Post
    ...that can impact reproduction include embryonic lethality, a short lifespan, or infertility and can arise...modeling blog series, we covered techniques that can be used to introduce genetic modifications into mouse...growing colony of genetically engineered mice, what can you do? In this post, we’ll cover why and how to ...transgene in order to see an effect. Hemizygous mice can be included in breeding strategies identical to those...mutation at one, but not both, of the alleles. You can cross homozygous or heterozygous mice from your F1... of integration of the transgene. These problems can be mitigated in different ways. Prior to engineering...transgene. In existing strains of mice, the transgene can be maintained on a hemizygous or heterozygous background...

  6. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    ...Overexpression methods flood the cell with protein, which can both interfere with a cell’s normal function and ...to ensure the edited cells are behaving normally can be prohibitively expensive for many labs. We’ve created...information and images generated from these cell lines can be found in the Cell Catalog on our data portal. ...components to physically enter the cells so they can go to work. Figure 3: Inducing the double ...Cell Catalog on the Allen Cell Explorer, where you can also find the certificate of analysis provided with... The editing strategy and design described above can be used to create similar donor plasmids for introducing...

  7. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    ...your experimental RNA. To solve this problem, gRNAs can be chemically modified to resist degradation while... Biotechnol; Rozners, JACS). These modifications can be done in-lab using commercially available kits ...surprisingly well-tolerated by the Cas9 protein and can increase on-target efficiency. In the same vein, ..., locked nucleic acids or bridged nucleic acids can also be introduced in the gRNA. These modifications... 20-bp targeting region of the gRNA. These gRNAs can be cleaved and rendered useless after less than a... the gRNA is released and the CRISPR-Cas9 system can initiate targeted editing events. Both technologies...There are many suitable dyes to choose from and you can opt to do-it-yourself with a kit or purchase it commercially...

  8. GPCRs: How Do They Work and How Do We Study Them?

    Type
    Blog Post
    ... subtype in the brain that can either activate or inhibit neurons. You can find information more about...inflammation, and neurotransmission. These receptors can be activated by an array of different ligands, including...signaling  So with the hundreds of GPCRs out there, how can scientists decipher the signaling pathways from ligand...the transcription factor from the receptor, which can then activate expression of a luciferase reporter...potential to discover new ligands for unknown GPCRs. You can find the entire PRESTO-TANGO kit at Addgene! TRUPATH...the 16 non-visual heterotrimeric G proteins that can be activated downstream of GPCRs. This collection...subunits of the heterotrimeric G protein complex that can be used to detect G protein activation via bioluminescence...

  9. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

    Type
    Blog Post
    ...is highly flexible and can accommodate any genetic elements of interest. It can thus be tailored to address... Alan Wong. The complexity of biological systems can hinder our attempts to study and engineer them, but...specific genetic perturbations. The ease with which one can design and synthesize CRISPR guide RNAs (gRNAs) for...provides a means to create barcoded gRNA libraries that can be used to combinatorially modify the genome, screen...different gRNAs in each plasmid. Each gRNA combination can be tracked and quantitatively analyzed by sequencing... With a list of gRNA targeting sequences, you can then readily generate your barcoded gRNA library ...interrogate and understand complex biological systems. He can be contacted at [email protected].   References 1. Cheng...

  10. Experimenting in an Open Source lab: from CRISPR to Cats

    Type
    Blog Post
    ...The computer scientists can get involved in modelling, the electronics folks can build a robot worm on ...meaning that we can obtain known mutants at $7.00 a strain. Makerspaces like ours can also make accounts...technologies and spaces where interested parties can do research. This interest manifested itself in the...on top of that model, and the biologists can get involved a in variety of aspects from the molecular biology...but that would be useful to our model  so that we can use CRISPR/cas9 to build the mutants, analyse their...intend to focus on motor neuron knockouts so that we can perturb factors related to worm motion. Working on... area, please stop by. And if not, I suspect you can find a makerspace somewhere near you. The place to...

  11. Is this the right place for me? 8 tactics for choosing a lab

    Type
    Blog Post
    ...lab alumni as you can or schedule video/calls if they are not local. Alumni meetings can be the most informative...lab such a big deal? It’s actually something you CAN choose and it will make a huge difference for your...right and responsibility to choose a lab where you can thrive and do your best work. This post is focused...choosing a lab, but almost all of these guidelines can apply to any workplace or job. It amazes me when ...questions in the sections below. Mentoring relationships can help identify your priorities and needs, but sometimes...informative way of gathering data. Former lab members can give open, revealing information about the culture...opportunities with the lab head. Former lab members can give you a fuller picture. Consider talking to people...

  12. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ...so much for how bright you can make a reaction but for how sensitive you can make a reaction. NLuc requires...fluorescent proteins (FPs) can be problematic because excitation light can cause phototoxicity, autofluorescence...tools are fantastic examples of technologies that can accelerate your research. The creation of NanoLuc...by external light sources. These external sources can generate autofluorescence and have limited penetration...protein fusion (OgNLuc). Both fluorescent proteins can be excited by Nluc emission and the lab has dubbed...nm – a clear demonstration of why a BRET reporter can improve deep tissue imaging. The constructs for GpNLuc.... The technique of coupling FPs with Luciferases can overcome these problems but has been hampered by ...

  13. Antibodies 101: Validation

    Type
    Blog Post
    ...sensitivity, which can be problematic if you are trying to show that your antibody can distinguish between...results so that others in the community can benefit and we can all help improve antibody reproducibility...antibody has been appropriately validated so that you can have confidence in your results. But what is appropriately...that you receive; citing existing validation data can be acceptable. However, it is your responsibility...for the target antigen - Just because an antibody can bind to its target, does not mean that is all it ...represent five experimental strategies researchers can use to show that their antibody performs as expected... types are amenable to genetic manipulation, nor can all genes be easily knocked down or out.   Example...

  14. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    ...nuclease, can be used in conjunction with Cas13 to amplify the signal of a single target. Csm6 can cleave...laboratories. In the case of the COVID-19 pandemic, qPCR can be used to diagnose the presence of SARS-CoV-2 RNA...reporter. The detectable signal from this reporter can be measured and quantified to determine the presence...an RNA guided RNase (Abudayyeh et al 2016). Cas13 can be guided by a single CRISPR RNA (crRNA) to cleave...non-specific ssRNA cleavage. How does SHERLOCK work? Cas13 can be programmed with crRNA to target an ssRNA of interest...recognizes and binds to the programmed sequence, it can promiscuously cleave surrounding ssRNA molecules....speed, ease of use, and portability. This method can be used for an array of applications including clinical...

  15. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

    Type
    Blog Post
    ...not all cell types can be labeled, and that while multiple copies of the transgene can integrate into cells...recombination is required before fluorescence can occur. Cre recombinase can be used alongside common, existing ...label an organ or entire organism. Because each cell can have multiple copies of the transgene that will recombine...express the construct but do not undergo recombination can be detected by immunostaining of the truncated YFP...and fluoresce in the same color as the fused XFP can be used to amplify the signal. Imaging endogenous... extremely versatile technology as recombination can be induced in developing fish via Cre protein microinjection...mouse (Snippert et al., 2010), a reporter mouse that can be used to track any cell type when crossed with ...

  16. Plasmids 101: Inducible Promoters

    Type
    Blog Post
    ...protein cannot bind. After an inducer binds to the activator protein, the activator protein can bind to...this post, we’ll discuss inducible promoters, which can be switched from an OFF to an ON state, and how you...inducible promoters regulated? Inducible promoters can be regulated by positive or negative control. Positive...temperature, and light are all examples of factors that can lead to the induction of a promoter. Below, you’ll...cAMP complexed with cAMP activator protein (CAP) can also stimulate AraC binding to I1 and I2 sites. Supplementing...near-zero expression at regular temperatures but can be induced by heat or cold exposure. Examples include... pBad is likely a better option since expression can be repressed by glucose. Temperature inducible promoters...

  17. Flow Cytometry Readouts: Yes, No, and Everything in Between

    Type
    Blog Post
    ...may not be clearly picked up by the sensors. You can adjust this by changing the voltage for the particular...1). Based on where the GFP- population ends, you can draw a gate which selects anything to the right of...expression, instead of its presence or absence, the plots can look quite different. You might have high expression...upregulated or downregulated based on your experiment. You can see from Figure 2 that determining where to place...quantify smeary plots For smeary expressions, you can quantify shifts in signal using the mean fluorescence...peak that shifts to the left. When using MFI, you can still continue gating populations of interest with...expression to the baseline/negative control. You can see how it is easier to visualize these four populations...

  18. With Our New Viral Service, We're Taking CRISPR Further!

    Type
    Blog Post
    ... with large inserts can make lentivirus production frustrating - a researcher can complete an entire lentiviral... SpCas9 and Cas9 nickase for genome editing. You can also enhance expression of your gene of interest ...EGFP, BRAF, and MAP3K4. The gRNAs targeting EGFP can be used as positive controls for gene editing or ...repository are CRISPR pooled gRNA libraries that can be used to screen for genes involved in a wide variety...Getting these quality-controlled reagents from Addgene can greatly expedite a researcher's ability to make discoveries...you’ll also receive transfer plasmid DNA that you can use along with lentiviral packaging plasmids to create...standard lentivirus production protocol here and you can find more tips and protocols for working with lentivirus...

  19. A Novice's Perspective on Neuroscience - SfN2017

    Type
    Blog Post
    ...If there’s a plasmid or AAV tool that you can’t find, you can always suggest a plasmid to us, request that...including a team that created a novel form of AI that can learn new tasks and beat humans at the ancient Chinese...I met one researcher studying how the microbiome can affect Alzheimer's - a connection I wouldn’t have... get into the field, there’s likely some way you can use the tools and skills you’ve developed in your...maybe your favorite FRET-based biosensing technique can be applied to an important neuronal signalling pathway...ve got a useful tool, there’s likely someway you can apply it in neuroscience. AAV tools from Addgene ...useful, these vectors require a bit of expertise and can be difficult for individual labs to make efficiently...

  20. Plasmids 101: Methylation and Restriction Enzymes

    Type
    Blog Post
    ...modification systems, yet can still affect whether certain restriction enzymes can effectively cleave DNA...of mutated plasmids which can later be verified by Sanger sequencing. How can I tell if my enzyme will ... of the DNA surrounding your restriction site(s) can help you quickly predict which (if any) of those ...sequence visualization software (we used Snapgene here) can help you assess whether your restriction site will...in Lane 2. Controlling methylation Finally, you can control methylation by altering your choice of bacteria...that will be blocked by Dam or Dcm methylation, you can ensure this site remains unblocked by first cloning...
Showing: 381 - 400 of 1078 results