We narrowed to 765 results for: Mos
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TypeBlog Post...(although there are exceptions). For example, on most flow cytometers you will need compensation between...cytometer’s setup. For instance, BV510 and AmCyan have almost identical emission spectra but may be called BV510...become a perpendicular vertical line. In general, most spillover between samples can be fixed with fairly...required. In Figure 3, we also know that on most cytometers, FITC and APC should not require compensation...
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Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria
TypeBlog Post...right Tn7 end sequences. Donor bacteria with a chromosomal copy of the RP4 transfer machinery mobilize these...and recipient bacteria that lack an integrated chromosomal copy of Mobile-CRISPRi. For firmicute gram positive...tri-parental mating. Donor bacteria contain a chromosomal copy of the RP4 transfer machinery, which mobilizes...versus the number that grew on a non-selective plate. Most bacteria tested had 1 transfer in every ~100 to ... -
CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments
TypeBlog Post...gRNAs (CFD < 20) bind almost not at all, and strong gRNAs (CFD > 80) bind almost as much as perfect gRNAs...performs HDR. Optimal mutation in-depth One the most sophisticated features of CrispyCrunch is how it... it is below 0.03, it stops mutating. (Note: the most common synonym in the human genome is chosen for... -
PCR Applications for Copy Number Variation Assays
TypeBlog Post... Signaling molecules: Sybr Green and Taqman qPCR most often uses either Sybr Green or Taqman for signaling...background noise, due to the large amount of dsDNA in most starting materials, and is not particularly sensitive...rather than by comparison to a standard curve, and most results can be confidently interpreted as integers...experiment. You’ll have to consider: Resources needed Most molecular biology labs and departments will have... -
Transferable Skills: Negotiation
TypeBlog Post...loses. Although this approach to negotiation is the most straightforward, it can mean that everyone leaves... why? Do they have an important plans? What is most important to each person? Someone has to be the...because of the need to be thoughtful about what is most important to each person. It can especially be difficult...organizations that partner with Addgene. What are the most important issues for our partners and how can Addgene... -
Plasmids 101: 5 factors to help you choose the right cloning method
TypeBlog Post...should you use if the budget is particularly tight? Most cloning processes will require a PCR reaction or...high chance of success (>95% for a single insert), mostly due to the use of toxic ccdB to prevent any non-recombinant...goes on... In this post, we’ve reviewed many of the most popular cloning methods out there, and I hope we...could choose from to perform these experiments. In most cases, there will be more than one right answer,... -
Transferable Skills Guide: Managing a Team
TypeBlog Post...the professional career of. Like everyone else I mostly learned through experience and from my mistakes...PhD students and postdocs we are used to doing (almost) everything ourselves and it’s a hard habit to ...done and how to prioritize but I just added the “most important” tasks my schedule. This was bad for 2...work, set expectations, answer any questions, and (most importantly) decide on a timeline. My experience... -
Experimenting with New Careers while in Grad School
TypeBlog Post... an awareness and understanding of science. Like most science PhD programs, mine was geared toward careers...along the way and hope they’ll help you make the most of the opportunities you’re provided throughout ...the State Museum has been the best supporter and most productive partnership CINSO has had. Volunteer!...happy. I started volunteering in the “Dinosphere” almost every week. I gained experience explaining science... -
Anatomy of a Plasmid Page at Addgene
TypeBlog Post...information on our plasmid pages? Well, to help make the most of these pages, we've written this post to guide... (kit, pooled library, etc) will be sent to you. Most of Addgene’s single plasmids are sent as bacterial... with a tag, it’s terminus will be listed as the most likely terminus the tag will end up on after successful... are listed, as well as the name of the strain. Most Addgene plasmids are maintained in the E. coli lab... -
Summer SciComm Series: Modes of Communication
TypeBlog Post... of options and then pick whichever is most accessible or most appealing to you. And if you’re not sure...users readily accessible via web searches; likewise, most in-person events and venues will have a solid understanding... math probably didn't sound like a great idea to most people in 2005, but XKCD is pretty successful! ... -
CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics
TypeBlog Post...human genome as a map with three coordinates: chromosome, start, and stop. For instance (chr3, 1, 10) ...stretch of DNA at the very beginning of the third chromosome, ten base pairs in length. An emerging family...types from the ENCODE Project. DHS sites are the most inclusive markers of regulatory regions in the genome...700 terabytes per gram—and form the X-shaped chromosomes we all know and love. To understand compaction... -
Fluorescent Proteins 101: Green Fluorescent Protein (GFP)
TypeBlog Post...Tyr66, and Gly67). When first discovered, one of the most surprising aspects of GFP was the fact that the ... importance of the entire GFP protein, which is almost completely required to maintain fluorescent activity...to increase its range of color and application Almost as soon as its sequence was elucidated, scientists...constantly being developed! Fusion tagging: One of the most common uses, GFP can be fused to the N- or C-terminus... -
Plasmids 101: Shuttle Vectors
TypeBlog Post...vectors in fast-growing bacteria first. In fact, most yeast vectors and many mammalian vectors are designed...will be lost from the population. A bacterial chromosome has a single origin of replication (ori), where... the cells that contain it. Antibiotic selection Most bacterial plasmids used in the lab contain an antibiotic...gene for growth in bacteria. Expression You’re almost there: you’ve planned for vector replication and... -
Twenty Years of Sharing: Expanding Accessibility
TypeBlog Post...material is right for their experiment. Table 1: Most popular guides, protocols, and collections of 2023...visitors in 218 countries. In Table 1, you can see our most popular educational resources for each type of resource...different countries (Figure 4). Four of the five most popular videos are protocol videos, where showing...a step can provide needed clarity (Table 1). Our most popular video, however, is “What is a Plasmid?”,... -
Viral Vectors 101: Preparing Pooled Libraries
TypeBlog Post...distribute several of our most popular CRISPR pooled libraries and one of our most popular barcoding libraries... cDNA, barcoding, screening, and gRNA libraries. Most of the pooled libraries at Addgene are lentiviral...thaw cycles, we include a 1.3 mL aliquot with mostlibrary requests specifically to allow scientists to ... -
Supporting Reproducibility with a Connected ELN
TypeBlog Post... for reproducibility than others. Unfortunately, most ELNs are designed as closed ecosystems and act as...graphic depicts, RSpace supports linking to all of the most commonly used online file storage tools, as illustrated...depicted in the graphic, RSpace is integrated with most of the tools noted above, and has a modern API. ... used and/or referenced in the experiment. Since most ELNs, unlike data repositories, are designed to ... -
New CRISPR Tools: Cas7-11 and PASTE
TypeBlog Post...off-target breaks, almost always resolved without any toxic effects to the cell. This means almost no indels and...retargeted. This finding was surprising – in the most common RNA targeting system, Cas13, the "collateral...one-size-fits-all approach to gene replacement therapy for most patients. But how does one go from “cut and paste... -
Antibodies 101: Immunoprecipitation
TypeBlog Post...protein is then eluted. This method is appropriate for most situations. A slightly different method is desirable...antibody to bind to the target protein. One of the most common immobilization methods utilizes bacterial...Consequently, users need to choose the protein type that is most compatible with the host species and isotype of ...Protein A/G, was developed, which can be used for most antibodies compatible with either Protein A or G... -
Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor
TypeBlog Post...of the mouse chromosome. Of the genes located in this stretch of DNA, Ly6a seemed the most likely candidate...SNPs and small insertions and deletions (indels) most likely to have large phenotypic effects such as ...fails to upregulate transduction efficiency in the marmoset brain." Neuroscience letters 665 (2018): 182-188... -
pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs
TypeBlog Post...non-integrating RNA virus Episomal vectors- extrachromosomally maintained DNA plasmids that express reprogramming...been used successfully to derive iPSCs from PBMCs. Most laboratories use Sendai virus to reprogram PBMCs...encoding reprogramming factors behave as separate extrachromosomal elements in the nucleus of the target cell...colonies are obtained by 10 days after electroporation, most of which are integration-free, karyotypically normal...