...characterized orthologs from “Anaerobic digester metagenome” (AdmCas13d) and Ruminococcus flavefaciens (nicknamed...87% mCherry protein knockdown measured by flow cytometry, respectively. Cas13d CRISPR array processing ... al. compared CasRx to multiple RNA regulating methods: small hairpin RNA interference, dCas9-mediated...

... can change depending on the location. Current methods for visualizing the location and function of microbes...reporters have limited deep-tissue performance, and sometimes require an alternative such as a radioactive tracer...and create ultrasound contrast. At a very low volumetric density and with a resolution of less than 100...

... a look at a few screening methods. Blue-white screen A widely used method of screening is the blue-white...sequencing primers in our Molecular Biology Reference. Sometimes common  primers can't provide all of the information...

...variety of cloning techniques and troubleshooting methodologies drew me to learn more about the process that...immunofluorescence-immunocytochemistry (IF-ICC.) This method was chosen because cells transfected with a plasmid...available from each plasmid webpage. Importantly, the methods developed by the Trimmer Lab for construction and...

...Clouse While many CRISPR-based genetic screening methods have primarily used Cas9 to knock out genes, researchers... screening toolkit by optimizing a Cas12a-based method for screens. Cas12a’s ability to perform multiplex...in parallel. Lastly, this Cas12a-based screening method was used to create a genome-wide library capable...

...transcription factors, starting with an established method called ChIP-seq. ChIP-seq relies on antibodies ... hundreds of transcription factors. We wanted something that would allow automation. This design cuts ...pressure on many other labs that may be doing something similar. There may be other variants of this that...

...subgroup of researchers in your area of expertise? Sometimes, the funders of your research may stipulate that...models, and software tools in the Materials and methods section of their manuscript to make resources easier...information you provide on data, materials, and methods, the more valuable your paper will be to the scientific...

...to understand the basis for cardiovascular and metabolic human diseases. They do that by studying patients...number of genes involved in cardiovascular and metabolic diseases. We are knocking out genes that have ...to examine the effect of a gene on cholesterol metabolism. The TALEN kit is quite useful in generating ...

...super-resolution microscopy. Cell Rep Methods 2(10):100301. https://doi.org/10.1016/j.crmeth.2022.100301  Additional...affinity reagents, and for good reason. However, sometimes you want a tool that is a little more compact ...

...panicked silence, I opened my mouth to panic-babble something about Western blots that made no sense, even in...options were to stop talking until I could present something coherent or to quickly string together all the... above to make a strategy that can help. Even something as simple as telling yourself, “I can stop and...

...pharmacological or genetic approaches, but only a few such methods exist and they only work for a subset of kinases...kinases. The Problem:  There is no common method for creating optically controlled kinases. The Solution:...KTR-mRuby2 system has several advantages over existing methods. First, it avoids activating other signaling pathways...

...efficiency was similar to that of a comparable FISH method. Importantly, they did not observe labeling when...with both intronic and exonic gRNAs. In fact, the method is specific and sensitive enough to detect gene...genome editing, activation and repression.” Nat Methods. 12(11) (2015):1051-4. PubMed PMID: 26344044. PubMed...

...is useful for many metabolic engineering and synthetic biology goals. In metabolic engineering, the expression...or can interfere with the precision needed in metabolic engineering or other synthetic biology circuits...

...enzymes that you are using are not blocked by methylation like XbaI, ClaI, etc. If you are doing multiple...to be strategy specific, there are few general parameters that you can adjust: temperature and time of ...molar ratio of 1:2 (plasmid:insert), but this parameter has to be optimized according to the strategy ...

...CASANOVA plasmids here! Bubeck F et al., Nature Methods 2018. PubMed PMID: 30377362. Studying neuronal...Energy Transfer (FRET, Heckler et al., 2015) or ratiometric Bimolecular Fluorescence Complementation (rBiFC...2in1 plasmid kits at Addgene! Mehlhorn DG et al, Methods Mol Biol, 2018. PubMed PMID: 29043675.Grefen C ...

...visualized with fluorescence or immunohistochemistry methods, these proteins are routinely fused to ‘tag’ or...techniques. Thus, these proteins provide a versatile method to identify the expression of transgenes of interest...Saccharomyces cerevisiae by use of an epitope addition method. Mol Cell Biol 8:2159–2165 . https://doi.org/10.1128...

...diversity and flexibility of diabodies. Methods. 10.1016/j.ymeth.2018.09.005 Muyldermans S. (2013). Nanobodies...including proteins, peptides, amino acids, drugs, metal ions, and cells. They can be used either with or...

...been “working on new experimental and theoretical methods to push the scale of genetic engineering, with ...carry CRISPR circuitry, it can slow their growth. Something has to be done about that.  The second problem...variation. The first one worked in each case. Sometimes in CRISPR people talk about off-target interactions...

...coaching plans that are financially accessible. Sometimes only a few hours of coaching time can make an ...Coaching. One nice thing about a group is that sometimes this offers the option of matching with the best...a good match as a coach for many scientists.  Sometimes it can be helpful having a coach that can offer...

...minute! Fun fact! Electroporation is one popular method scientists use to promote transformation, but it...offer resistance to toxic substances like heavy metals or pesticides, enabling their hosts to survive ...inducible expression systems that allow tuning of metabolic pathways. Fertility (F) plasmids: microbial sharing...