We narrowed to 993 results for: Des
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TypeBlog Post...semester one and then proceed to use those skills to design and carry out their own novel experiments in semester...resources, offers mentoring for instructors, and provides opportunities for students to exchange ideas and...results in student learning gains and positive STEM attitudes in CUREs. CBE Life Sciences Education. 21(4):ar74...
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Plasmids 101: Secondary Nanobody Toolbox
TypeBlog Post...their production requires animal sacrifice and precludes genetic engineering of a tag or fusion with a ... with secondary nanobodies conjugated with the desired fluorophores. In figure 4B below, triple colocalization...review. If you just want to use the nanobodies described in Pleiner et al, the key steps for generating... -
MXS Chaining
TypeBlog Post...Origin and purpose of MXS chaining MXS-chaining was designed to create plasmids for fluorescence imaging or... can be assembled. For example, because CpG dinucleotides are underrepresented in most vertebrate genomes...and MluI recognition sites, which contain CpG dinucleotides in their 6-mer sequences, are rarely found in... -
Plasmids 101: Protein tags
TypeBlog Post... Protein tags are usually smallish peptides incorporated into a translated protein. As depicted in the...depending on their protein of interest. After several decades of trying to address these challenges, researchers...examples. The tables list more common tags than are described in the post, but have been categorized to help... -
Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More
TypeBlog Post...worked extensively with enChIP systems. Can you describe this technology and its advantages? Fujii: In ...enChIP system using CRISPR technology. Can you describe this technology and some of its potential applications... any other technologies that you would like to describe? Fujii: I believe that "Biology is Method", i.e... -
Fluorescent Proteins 101: Green Fluorescent Protein (GFP)
TypeBlog Post...biochemical properties. In 1995, Roger Y. Tsien described an S65T point mutation that increased the fluorescence... in the wildtype protein and facilitating its widespread use in research. Many other mutations have since...array of GFP-based fluorescent biosensors has been designed to detect a variety of intracellular conditions... -
Hot Plasmids: Fall 2024
TypeBlog Post...promising tool for future structure-guided drug design and studies of protein ubiquitination and degradation...CRBNmidi plasmid here! Kroupova, A. et al. (2024). Design of a Cereblon construct for crystallographic and...specifically for C. elegans (Cao et al., 2024). The team designed coding sequences for mScarlet3 and mScarlet-I3... -
To Codon Optimize or Not: That is the Question
TypeBlog Post...Usage Tabulated from GenBank database (CUTG), that provides an electronic dataset for codon-usage of 257,468...(A) The codon wheel depicts the 64 codons that encodes the 20 amino acids and three stop codons. (B) Example...Codon Optimization ATUM DNA2.0 IDT Synthetic gene design made easy GenScript- GenSmart Codon Optimization... -
Antibodies 101: Immunoprecipitation
TypeBlog Post...most situations. A slightly different method is desirable if the antibody is expected to bind weakly to ...vary in shape and structure. Their porous surface provides a large area for binding, but antibodies conjugated... slightly higher, than those of agarose beads, despite the size difference. Magnetic beads are particularly... -
CRISPR-based Genome Editing Systems For Editing in E. coli
TypeBlog Post...CRISPR-associated transposases” or MUCICAT. Here, we’ll describe each of these CRISPR systems. The original pCas...by sacB counterselection. The MUCICAT toolkit includes versions of plasmids with different induction ...mechanisms behind their distinct phenotypes and to design a new generation of engineered cells for green ... -
Antibodies 101: Reading a Flow Plot
TypeBlog Post...series of markers are set up sequentially, from broadest to most specific marker, and are represented as...are, try drawing a quadrant over the plot that divides it into four, using the location of the cell populations...Gating and plot orders As the gates progress from broadest markers to most specific, the first step is to... -
Viral Vectors 101: Preparing Pooled Libraries
TypeBlog Post... If not, our protocol on Lentivirus Production provides a general approach for producing lentivirus in...excellent protocol paper by the Feng Zhang lab describing construction, lentiviral production, transduction...the Broad Institute- A variety of protocols and guides regarding pooled libraries, from cloning a pooled... -
New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression
TypeBlog Post...to distinguish from background tissue unless destructive ultrasound imaging was used. We set out to develop...acoustic properties of these gas vesicles made non-destructive specific imaging in tissue difficult. The first-generation... allowed the GVs to be flexible and produce nondestructive nonlinear ultrasound contrast. It turned out... -
Important Considerations in Optogenetics Behavioral Experiments
TypeBlog Post...other words, your optogenetic experiment should be designed around a behavioral test in which your circuit...of mouse transgenics. Everything that has been described is intended for the mouse; however, all the steps....Diester, Ilka, et al. "An optogenetic toolbox designed for primates." Nature neuroscience 14.3 (2011)... -
Adeno-associated Viruses (AAVs) for Genome Editing
TypeBlog Post...human cells (Kim et al, 2016). The Waldman lab has designed their AAV tagging vectors (pAAV-SEPT-Acceptor ...vectors for your specific gene, the lab recommends designing homology arms with the modification of interest...combining the AAV and CRISPR-based approaches is described in several recent reviews (Howes and Schofield... -
CRISPR 101: Off-Target Effects
TypeBlog Post..., it’s essential to understand them in order to design the most specific assays possible. In this blog...-target effects should impact your experimental design differently. Yes, it is always better to reduce...introducing a specified edit). However, if your design does have wiggle room, you can minimize off targets... -
PiggyBac-ing Through the Genome Editing Field
TypeBlog Post...facilitate the selection of cells containing your desired edit. The selection is marker is then seamlessly...sites is cytotoxic for proliferating cells and provides a simple tool for conditional cell ablation.,”...Zinc-finger nuclease-induced gene repair with oligodeoxynucleotides: wanted and unwanted target locus modifications... -
A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression
TypeBlog Post...experiments to minimize it. Overview of DIO/FLEx vector design Recombinase-dependent vectors utilize pairs of ...between them. Early recombinase-dependent vector designs used a lox-STOP-lox (LSL) or frt-STOP-frt (FSF)...you intend to use performs in your system. This includes a thorough testing of both the serotype and promoter... -
Sequencing Options for CRISPR Genotyping
TypeBlog Post...good starting point for most researchers as it provides a list of putative off-target sites that they ... approach misses larger mismatches (e.g. six nucleotides) that may still lead to off-target double-stranded...contains both the rare off target event and the desired edit is low. Therefore, unbiased sequencing may... -
Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins
TypeBlog Post...tetroxide). These conditions destroy most PA-FPs, but, in 2015, the Looger Lab designed 2 variants of EosFP that...to green). For example, mEosFP (named after the goddess of dawn in Greek mythology) switches from green...