We narrowed to 904 results for: CAN

...they need to do before they can be good at it. Like any other skill, it can be developed with education...convert to the principle that you can't motivate people to do things, you can only demotivate them. The primary...if you are good at science and you are smart, you can be a good manager. The kind of smarts and the type...above – what type of bad manager have you had?). You can avoid some of these common pitfalls of management... an extreme by scientists) Are highly mobile and can move to a new workplace if opportunities for learning...primary obstacle of job dissatisfaction, the work can get done, and get done well. The rest of the blogs...series will address more specific tactics managers can utilize to be in touch with the needs of the team...

...screening project can be a tremendous undertaking, producing a wall of results that can only be described...kinome is a well-annotated collection of genes that can provide a useful starting point for developing a ...two sets, each with 4 gRNAs per kinase, so screens can be performed with either 4 or 8 gRNAs per gene. Further...more gRNAs found in genome-wide libraries, and thus can be screened in pooled format at a more-typical scale...plasmid formats A first for a library at Addgene, you can also order the individual lentiGuide gRNA plasmids...individual plasmids separately. Additionally, this set can be used for cherry-picking individual gRNAs that ...

...method that can handle all applications of iPSCs, but there should be at least one method that can help you...transgene-free and vector-free cells No genomic integration Can be difficult to fully remove virus from cells Plasmid... many variables (cell type, criteria used, etc.) can effect reprogramming rates and such a number maybe...also creates heterogeneous iPSC cell lines, which can complicate comparisons made between lines. Incomplete...polycistronic vector or multiple single-gene vectors can be used to package reprogramming factors. An important...re-expression of viral transgenes in iPSC-derived cells can interfere with their differentiation potential. Finally... as well repressed following reprogramming. This can be overcome by using a Tet-inducible virus to allow...

...the potential CRISPR technology can have on the human microbiome. You can consider the microbiome as a collection...selection step for successful modifications, efficiency can be low, especially for larger modifications. What.... The system also functions in S. pneumoniae and can be used to generate multiple mutations simultaneously...takes two days, and the pTargetF and pCas plasmids can be subsequently cured from the bacteria. Although...using BbsI and Golden Gate Assembly. Either plasmid can be also linearized with XbaI to insert extra elements... a multi-spacer CRISPR array, CRISPR-transposons can be multiplexed. Transcriptional repression (CRISPRi...dead (dCas9) targeted to a promoter or gene body can repress transcription by physically blocking the ...

...color emission. As you can see in Figure 1, emission spectra of neighboring colors can and do overlap. For...and the markers they label) can occur. As I mentioned previously, you can also use fluorescent reporters... inside of your cells. While proteins themselves can emit intrinsic fluorescence when excited with ultraviolet...only occur in combination with that reporter, you can infer its presence through detecting the reporter's...combinations for different markers of interest, you can separately detect these markers in your analysis.... photons excite electrons of a fluorophore, they can do so at a range of wavelengths, rather than at just...names, for example, BV421 and RB545. Fluorophores can also come in combination of two conjugated single...

...expressed genes can be the next stage in making sense of such data. These targeted screens can allow a researcher...tools to study the immune system and cancer. More information can be found through https://wermelinglab.com...Medicine, Solna, Karolinska Institutet, in Sweden. It can be very time consuming to design 5 guide RNAs (gRNAs...CRISPR screen. Luckily, the Green Listed software can help you do just this, probably in less than a minute...properties, migration abilities, or other phenotypes that can be efficiently measured at the single cell level)...receptors expressed on the cell surface. Thus, a screen can be used to target surface proteins to determine which.... Once you have your list of genes, Green Listed can help make the gRNA design process quick and easy....

...The localization of the fusion protein can be verified and it can be compared to what is expected based...catalytic activity or interactions) can be challenging. If a fusion can complement a knock-down or knock-...the chimera are encoded by DNA. Since researchers can generate almost any DNA sequence in the way that ...the native properties of the protein of interest can be challenging. In this blog I discuss strategies...derived from Aequorea victoria GFP sticks out and can be considered a linker (figure 1). In fact, for several... and N-terminus of the donor fluorescent protein can be truncated to increase the FRET efficiency. Creating...fusion,” but I find this very confusing because it can mean that the fluorescent protein is at the C-terminus...

...the candidate off-target sites. For example, if there are 10 putative off-target locations you can 1) ...sequencing methods. Candidate site sequencing Sites with high sequence similarity to the gRNA can be sequenced...sequenced. These sites can be selectively sequenced to determine if there are significant editing events, serving.... In this blog, we will discuss when off-targets can be an issue in CRISPR, how to predict and minimize... nucleases are promiscuous. In other words, they can sometimes localize to unintended sites with sequence...reduce off-targets and patient risk. Hopefully you can understand that depending on your platform, off-target...predicted frequency of off-target events, many of which can be combined to lower them even further. While not...

...thereby generating a complete, circular plasmid that can be easily maintained in a variety of biological systems...proper orientation downstream of the promoter, you can add an EcoRI site just 5’ of the start of the gene...site just 3’ of the end of the gene. This way you can then cut the plasmid backbone as well as the insert...experiments.     Alternatively, this whole process can be completed using a single enzyme if your insert... that enzyme’s restriction sites, but the insert can then anneal to the backbone in either a forward or...Many companies now sell fast digest enzymes that can digest large amounts of DNA in as little as 10 minutes...migrate slower than smaller linearized fragments. You can separate your backbone away from any inserts cut ...

...high accuracy. Conversely, you can only run one sample at a time, which can become costly and impractical...is generally cheaper if you have many samples and can output more data, and it typically results in high...structures, or repetitive regions. Short-read sequencing can also be time-consuming, depending on the library ... to hundreds of base pairs, long-read sequencing can read sequence lengths in the thousands or even millions...by the instrument changes. Base-calling software can then identify the nucleotide that passed through ...technology. Created with BioRender.com. Nanopore can provide longer read lengths of 10 kb to 100 kb (or... to sequence an entire plasmid in one read. Runs can be completed in anywhere from under an hour to 72...