We narrowed to 709 results for: esc
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TypeProtocol... several μg/μL. The protocol below is meant to describe the general procedure for purifying plasmid DNA...
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What is Polymerase Chain Reaction (PCR)
TypeProtocol...Description of Polymerase Chain Reaction with protocol, tips and FAQ... -
What's New in CRISPR - March 2020
TypeBlog Post... in yeast Base editing CRISPRi in Caulobacter crescentus Multiplexed gRNA for human pluripotent stem cells... base editor plasmids CRISPRi in Caulobacter crescentus Figure 1: Serial dilutions of CRISPRi in...adapted from Guzzo et al., 2020. Caulobacter crescentus is a popular model organism for studying the ...stem cells Lindy Barrett’s lab developed new fluorescently labeled piggyBac vectors to deliver uniform ... lines with AAVS1-integrated, inducible, and fluorescent dCas9-KRAB and dCas9-VPR for quantifying and ...that express both the dCas9 construct and the fluorescently-labeled gRNA multiplex. Read the paper in Scientific... -
Zebrafish as a Model for Behavior: Swimming into the Optogenetic Spotlight
TypeBlog Post... and allow for noninvasive live imaging with fluorescent tags and activation of light sensing proteins...distinct behaviors such as spontaneous swimming and escape reflex. These traits, coupled with short generation...zebrafish at 5 days post fertilization, including escape behavior and spontaneous swimming. When startled...zebrafish react by performing a distinct C-bend to escape possible danger (see figure). Larvae also perform...in response to any specific stimulus. While the escape response neural pathway is well characterized, ... kind of swimming behavior, spontaneous swim or escape, was elicited when these different neuron groups... -
RUBY-Red Siliques
TypeBlog Post...categories: antibiotic/herbicide resistance and fluorescent proteins. Selection using resistance genes usually...would only then kill the desired plants! PCR or fluorescent markers are currently used in these situations...from heterozygous individuals. Only 1/4 of the descendants of the heterozygous plants, therefore, are useful... I can do this by expressing a gene that rescues the mutations, using RUBY as a selection marker...mutation but lacking both RUBY and the construct that rescues the mutation. I will be able to easily choose the...could be achieved much easier than by PCR or a fluorescent microscope. First, red CRISPR-positive transformants... -
RANbodies: Reporter Nanobody Fusions
TypeBlog Post...and instead are conjugated to reporters (i.e. fluorescent dyes, HRP, etc. ) after purification. Antibodies...signal amplification occurs when HRP converts a fluorescently labeled tyramide molecule into a highly reactive...linked to residues within close proximity thus fluorescently labeling them. HRP RANbody staining results ... ≥100-fold increase in signal over intrinsic fluorescence. Increased signal is likely due to lower background...and tissue sections Amplifying weak signals of fluorescent protein (i.e. GFP, RFP) Detecting GFP reconstitution... from Penn State Need help picking the right fluorescence microscopy technique to use? Check out this ... -
Virus Protocol - Generating Stable Cell Lines
TypeProtocol...from Feng Zhang (Addgene plasmid #52962 ) and is described in Improved vectors and genome-wide libraries ... -
AAV Titration by qPCR Using SYBR Green Technology
TypeProtocol...typically ROX dye), to normalize non-PCR–related fluorescence fluctuations and to minimize well-to-well variability... -
Hot Plasmids and Viral Preps - January 2021
TypeBlog Post... in bacteria CRISPR-Casɸ mGold, a new yellow fluorescent protein New CRISPR plasmids New items from the...32675376/ mGold, the most photostable yellow fluorescent protein to date By Justin Ng High-throughput ...High-throughput screening methods, such as fluorescence-activated cell sorting (FACS) helps researchers sift through...million cell types expressing variants of Yellow Fluorescent Protein (YFP). The team identified a new YFP ...Resources page. CDDR (CRISPR–Cas9-based Dual-fluorescent DSB Repair) is a double-strand break repair assay...the introduction & resolution of two DSBs in a fluorescent reporter. Find the plasmids here Yongsub Kim... -
Plasmids 101: FLEx Vectors
TypeBlog Post...study a wide variety of in vivo conditional gene rescue experiments, as well as for improvement of optogenetic...additional uses for FLEx switches: Conditional rescue of a gene knockout: To test if the phenotype you...by coding sequence of the gene you would like to rescue with in the antisense orientation (let’s call the... to test whether or not expressing GeneR alone rescues the phenotype being studied. Introduction of ...functional mutation in vivo: Similar to a conditional rescue experiment with a wildtype copy of a gene, you ...presence of Cre, GFP expression is lost, allowing for rescued expression of GeneR. Sometimes also referred... -
Anatomy of a Plasmid Page at Addgene
TypeBlog Post...plasmid, how will it be sent to me? Description: This describes how the plasmid or other product (kit...useful to me? Plasmid purpose: This is a brief description of what the plasmid was designed and used for...vector’s plasmid number here. Gene/insert: This describes what has been inserted into the backbone, and ... sequencing. Growth in Bacteria: This section describes how to grow the bacterial strain in which the ...associated, such as when a plasmid contains certain fluorescent proteins. Finally, for many plasmids you can ... -
Plasmids 101: Secondary Nanobody Toolbox
TypeBlog Post...Western blots. ELISAs. Immunofluorescence. What do all of these techniques have in common? They all typically... distance between the target antigen and the fluorescent label, which can reduce the potential image resolution... resolution of super-resolution fluorescence microscopy. Their size also limits their ability to penetrate...penetration and decreases the distance between a fluorescent tag and the target antigen to about 2 nm, which...review. If you just want to use the nanobodies described in Pleiner et al, the key steps for generating...: Purified nanobodies are then tagged with a fluorescent dye or HRP via maleimide-labeling. Maleimide-... -
Plasmids 101: Protein tags
TypeBlog Post...Which Fluorescent Protein Should I Use? Plasmids 101: Green Fluorescent Protein (GFP) Fluorescent Protein...Plasmids 101 has covered GFP and its related fluorescent proteins, which are sometimes used as tags for...LVPRGS Cleaves between Arg and Gly residues PreScission LEVLFQGP Cleaves between the Gln and Gly residues...examples. The tables list more common tags than are described in the post, but have been categorized to help... -
Finding nucleic acids with SHERLOCK and DETECTR
TypeBlog Post...quenched fluorescent ssRNA reporter is added to the reaction. Cleavage of the quenchable fluorescent RNA by...Cas13 and Cas12a, are combined with different fluorescent reporters for each enzyme. In addition, Csm6,...Csm6 and a Csm6 specific reporter (in the same fluorescent channel as the Cas13 reporter), they could significantly...papilloma virus (HPV) genome, via a crRNA. An ssDNA-fluorescently quenched (FQ) reporter, which will produce a...rate of ~1,250 cuts per second. A quantifiable fluorescent signal designates the presence of your DNA of... -
Tips for Writing a Good Cover Letter
TypeBlog Post...experience and skills would help you do the job as described. Let’s imagine you’re applying to a job that ... your cover letter, you can expand on that by describing how you developed and used a ddPCR assay to answer...start by looking at the requirements in the job description. For each requirement, ask yourself if you have...exactly what you would want to talk about! You could describe your organizational system, your rationale for...company/organization]. I am a [adjective that describes you] scientist with over [number] years of experience...the university]” Include keywords from the job description to help the hiring manager quickly make the connection... -
Bacterial Toxin-antitoxin Systems as Molecular Biology Tools
TypeBlog Post...plasmid addition was first described by studies of EcoRI and Bsp6I in Escherichia coli in 1995 when introduction...systems. As with the other toxin-antitoxin systems described above, one part of the pair is more stable than...toxin-antitoxin system discovered. Unlike the systems just described, the hok/sok systems consist of three components...proof of concept, they tested a lux-mediated bioluminescence reporter in vivo in the absence of selective..., Hanna Engelberg-Kulka, and Gad Glaser. "An Escherichia coli chromosomal" addiction module" regulated... -
Antibiotic Resistance: An Old Solution but a New Problem
TypeBlog Post...antibiotic’, very often we immediately imagine a prescription for a bacterial infection. While this post will...that the term ‘antibiotic’ can also be used to describe anti-parasitic drugs and anti-fungal drugs, because...development of resistance include (Figure 1): Over-prescription of antibiotics (for example, giving antibiotics...not finishing the whole course of antibiotics prescribed to them This may seem counter-intuitive considering...The take-home? Always take your medication as prescribed. How do we know which bacteria are resistant ...infection is still effectively treated. We can prescribe antibiotics for the shortest time period possible... -
Pouring LB Agar Plates
TypeProtocol...that way for ~10 min. This will allow any steam to escape from the autoclave and will cool your gel-mix slightly... -
Plasmid Cloning by PCR (with Protocols)
TypeProtocol...plasmid of interest. For this example, we will describe how to copy a cDNA from one vector into a new ... -
Coomassie Purity Stain of Recombinant Antibodies
TypeProtocol...Purification Protocol Introduction This protocol describes how to determine the purity and concentration ...