...to the popularity of GFP including (i) fast and complete maturation to functional, fluorescent protein ...diameter of about 2.4 nm (Hink et al., 2000). The complete beta-barrel is necessary for its fluorescence ...genetically encoded tag and exogenously added components. For a review see Lotze et al. (2016). To date...is used to excite GFP, may also excite several components that are naturally present in cells or media....

...lacY1 proA2 rpsL20 xyl-5 leu mtl-1 Stbl4   Electrocompetent cells for cloning and storage of unstable ...XL10 Gold Tetracycline and Chloramphenicol High competency cloning and propagation of large plasmids, ligated...tables are by no means exhaustive! For a more comprehensive list and additional information, visit OpenWetWare's...of Bacterial Expression Systems. Check out our companion post describing protein expression strains, where...

...certain Cas enzymes are less compatible with the bulkiness that accompanies mispairing of bases. To reduce...unnecessary for many applications, it is the only comprehensive method available. Targeted sequencing approaches...must be quantified slightly differently. For a comprehensive approach, RNA-seq would be an appropriate method...

...helps all viewers, especially if you have some complex patterns with several overlapping targets to interpret...you add the secondary antibody, it should get completely washed away and leave no signal. If you see lots...samples where you label only one target in each. Compare these individually-labeled samples with the results...deep phenotyping and spatial analysis of cells in complex tissues. Proceedings of the National Academy of...

...could avoid potential off target artifacts by comparing the phenotypes of multiple gRNAs. Targeting different...lanes 0.3 % 3.0, Figure 5, lane 2) is faint, so comparing it to the undigested control (Figure 4, lane 0...instead of agarose gives much sharper bands. I compared two MOIs of gRNA transduction to see if that had...testament to how far genome engineering has come. For comparison, in 2007 it took about 3 months to knockout a...

...your data by Nov. 12 to compete. The top ten participants will be invited to compete in our speed round data.... Bobae An will host a Q&A session after the competition, where you can get all your burning AAV questions...

...In complex metazoans, rapid cell division and large scale cell mobility are essential processes during... required for a growing organism to make the complicated transition from a clump of cells to a fully differentiated...organism. Studying cell mobility within these complex environments has never been easy and, for many ...labelling of cells using cre-lox is advantageous when compared to photoactivable or photoconvertible proteins...

...straightforward to learn and relatively cheap (compared to many other technologies and techniques in the...experiment work? What are the various tools and components necessary to run it? And, most importantly, what...writing. Derek is also a pianist and an aspiring composer. View his blog drsimonsaysscience.org or follow...applying optogenetic tools derived from direct comparative analysis of microbial opsins. Nature methods....

...fragments, allowing ordered assembly of multiple components into a destination vector. The first step in ...Oligonucleotides specifying the gRNA seed sequence should be compatible with type IIS enzyme BfuAI. Gateway cloning is...supply it with another plasmid.   Figure 3: Comparison of Multiplex Strategies including Standard PolIII-gRNA...Cas9-containing binary vectors. The system is compatible with both monocot and dicot plants. Liu Lab Golden...

...easier to generate mouse models.   Cpf1 Update: Comparison to Cas9 and NgAgo As you might expect given the...off-target editing and similar on-target editing when compared to the most commonly used Cas9 variant, SpCas9...Generating Mouse Models Using CRISPR/Cas9 Cpf1 Update: Comparison to Cas9 and NgAgo Special Delivery: Fluorophore...