...immunoprecipitation (locus-specific ChIP). These methods consist of insertional chromatin immunoprecipitation (iChIP...on development of technologies for biochemical analysis of genome functions such as transcription and ...technologies. The locus-specific ChIP technologies consisting of iChIP and enChIP are recent examples. We also...

...CRISPR corner New from the viral service Faster catalysis after yeast-based protein evolution By Joanne ...evolution platform to generate TEV protease variants. Catalysis by TEV protease allows a transcription factor ...CRISPR/Cas Toolkit for plants includes 95 plasmids consisting of CRISPR/Cas nucleases, base editors, gRNA backbones...

...singletons (reads that only appear once) in our analysis. Many pipelines toss these singletons out, and...stranger - Finding mentors for the meeting Just like analysis pipelines for DNA that group similar sequences...variability was organic, as some people need less assistance than others, but we did have some participants...

...the accompanying sequencing data collection and analysis eventually outpaced our software, even with continuous...singular purpose (ex. adapter trimming, quality score analysis, k-mer counting, or sequence assembly), but the...would handle plasmid sequences on a 96-well plate basis, but for rapid testing and refinements, we specified...

...concentration, ionic strength and pH, affect binding the lysis buffer may need to be adjusted depending on the ...include protease and phosphatase inhibitors in the lysis buffer, keep samples on ice, and perform the IP ...For mass spectrometry, elute in a urea-containing lysis buffer. Alternatively, the protein can also be eluted...

...pTargetF system Our original pCas/pTargetF system consists of one plasmid that expresses the S. pyogenes ...engineering case study of N-acetylglucosamine biosynthesis, we constructed a strain library with 1-11 copies... modified pCas/pTargetF system for CRISPR-Cas9-assisted genome editing in Escherichia coli. Acta Biochimica...

...important models for solutions to the coral reef crisis. I like how they are using protocols.io to create...disparate environments so can be used study the genetic basis of evolution in immunology, neurobiology, and more...lab has the capability) with receptor mediated endocytosis. They used this to edited those brown marmorated...

...They first created a defective chloramphenicol resistance gene (CamR) by introducing a point mutation (... adenine base editor would restore antibiotic resistance. To find such a protein, they created a mutant...any restriction enzyme (RE) recognition site, RE analysis of PCR product could be used to detect success...

... This is to avoid the chances of insertional mutagenesis, which is the integration of transgenes into ...insertion and which can potentially promote tumorigenesis in vivo. The phenomenon has been recorded in...Another drawback of using the Sendai virus is the persistence of residual viral material [6]. The complete ...

...Properties of FLP Recombinase Evolved by Cycling Mutagenesis.” Nature biotechnology 16(7): 657–62. PubMed ...Deleter Mouse in C57Bl/6 Based on FLPo Recombinase.” Genesis 48(8): 512–20. PubMed PMID: 20506501. 6. Logie,...Synapses Are Repeatedly Assembled and Removed at Persistent Sites In Vivo.” Neuron 89(4): 756–69. PubMed ...

...proteins within the bacterial genome. From this analysis, they identified 53 potential candidate genes ...within Cas13a are essential for RNA cleavage, consistent with known roles for HEPN domains in other proteins...located at the 3’ end of the spacer sequence and consists of a single A, U, or C base pair. In bacteria,...

... is the person you report or talk to on a daily basis who will help you get on your feet with techniques...accordingly in case you have questions or need a little assistance at some point during the procedure. Be an active...everything is magically made up by helpful TAs or assistants, and sitting out on the bench waiting for you...

...that a plasmid can transfer to, replicate in, and persist in. Some plasmids have a narrow host range, while...host range recombinant vectors Replicons, which consist of a plasmid’s ori and all of its control elements...scientists can incorporate interchangeable antibiotic resistance cassettes (Prior et al. 2010), inducible promoters...

...of its parts. Elements such as the antibiotic resistance gene and the origin of replication are necessary...elongation factor 1 alpha Constitutive  Tends to give consistent expression regardless of cell type or physiology...expression, but may vary by cell type. Tends to resist promoter down regulation due to methylation or ...

...ability to reassemble fragmented genomes is the basis of several split vector approaches developed in ...transgene expression from split vectors has been inconsistent. A greater understanding of basic AAV biology...vector strategies and lead to greater and more consistent transgene expression. References: Allocca M...

...promiscuous biotin ligase (Kim et al 2016). BioID2 consists of a humanized biotin ligase from A. aeolicus,... assemble when in close proximity. Split-BioID consists of a split BirA* molecule that can reassemble ...compared to the standard E. coli BirA* mutant. RapID consists of this mutant BirA* and an RNA component that...

...s group created Spinach2 using site-directed mutagenesis of Spinach. In doing so, they identified the ... IV, were then identified through microfluidic-assisted in vitro compartmentalisation for their high binding...PMC3852148. Warner, Katherine Deigan, et al. "Structural basis for activity of highly efficient RNA mimics of green...

... social science to an audience that I thought consisted of all biologists. The social scientists in the...to them with an answer, and do so if you can. 9. Resist the urge to say everything I allow 1 slide for ...topic or story is all you need, if you do it well.  Resist the urge to show all or even a lot of data.  I ...

...must be recoded to ‘TGA’ using site-directed mutagenesis. In the C321.ΔA strain, this TGA will be recognized... formation: the last puzzle of aminoacyl‐tRNA synthesis." FEBS letters 462.3 (1999): 302-306. PubMed PMID...Sauerwald, Anselm, et al. "RNA-dependent cysteine biosynthesis in archaea." Science 307.5717 (2005): 1969-1972...

...contributed by guest blogger Joachim Goedart, an assistant professor at the Section of Molecular Cytology...conditions needs to be examined on a case-by-case basis. GFP is sensitive to acid The chromophore of GFP...blogger, Joachim Goedhart! Joachim Goedhart is an assistant professor at the Section of Molecular Cytology...