...evolution of ligands by exponential enrichment (SELEX), some 1010 oligonucleotides could be screened for their...microarrays and biochips. Ultimately, there are just some applications that lend themselves to aptamer-based...Hanhui, et al. “Multicolor CRISPR labeling of chromosomal loci in human cells.” Proc. Natl. Acad. Sci. ...

... the excision of the phage from the bacterial chromosome and the regeneration of attP and attB sites. ...include the proper protein expression elements (ribosome recognition sequences, start codon, stop codons...attL2 only with attR2, and so on. Take a look at some of the Gateway Multisite plasmids available at Addgene...

...created a yeast-based directed evolution approach with some notable improvements. Directed evolution techniques...localization of biomolecules within cells. However, sometimes these fluorescent proteins might not localize ...collection to provide ready-to-use viral preps. Here are some of the new viral preps from recent months: New ...

...surface and a corresponding receptor in the host cell. Some of these envelope-receptor pairings are broad, allowing...by nonspecific complement mechanisms in serum. Sometimes this is due to the cells that the viral vectors... broad or specific to suit your needs. Check out some of these review articles to find tables and information...

...studying vertebrate embryogenesis. To illustrate some of the reasons that zebrafish have been embraced... generating mutants. Unexpectedly, phenotypes sometimes differ depending on whether a gene is deactivated... using lineage tracing in zebrafish To highlight some of the features that make zebrafish useful, I’ll...

...transformation process could save the plant biologist some time… months, in fact. In the Lee-Parsons lab, ...Mortensen et al., 2019). Our method might provide some useful guidelines for the development of transient... different treatments on transformation success. Some reporter gene examples are below. The GUS gene ...

... the search to a ~4.5 Mbp region of the mouse chromosome. Of the genes located in this stretch of DNA,...permissiveness While PHP.B has impressive CNS transduction in some strains of mice, these results are not reproduced...time AAV user tips from an AAV rookie  Learn about some considerations when using AAVs Learn more about ...

...ricin uptake. The ricin screen did fail to identify some known ricin-regulators, but most of these genes ...show that genome-wide CRISPR screens are prone to some of the same flaws as RNAi screens, such as non-specific...guides for off-target activity, Morgens et al drew some generalizable conclusions about off-target activity...

...use epitope tags to monitor single molecules - something researchers often want to do at neuronal synapses...and other FPs have the tendency to brightly label somata but not neuronal processes. In contrast, smFPs ...microscopy. Sample preparation for these techniques sometimes makes it more difficult to detect protein tags...

...surrounding cell culture over the last few decades some pharmacologists have tended to avoid cell culture... stored in liquid nitrogen long term, preferably some of these vials will be stored in a separate backup...changing but yet more is needed to convince people of some of the basic tenets of responsible cell culture ...

...difficult, and lacking in reproducibility. Read on for some tips for getting the most out of your lentiviral...virus is the second. When harvesting lentivirus, some labs prefer to collect and pool the harvests from...divided into single-use aliquots, and stored at -80°C. Some reports suggest rapidly freezing virus in a dry ...

...potential pitfalls of recombinant protein expression and some of the most popular commercial strains designed ...cell thus leading to slowed growth. Additionally, some protein products may be toxic to the host when expressed...within a reasonable time-frame. For these cases, some strains carry an additional measure of control such...

...Based on personal preference, some people like to make stricter gates while some like to make looser ones ... your cells on a flow plot, you might have to do some testing or ask experienced colleagues.   ...

...knock-ins Are you introducing a large insertion, something 20 base pairs or bigger? Then screening by PCR... pairs modified? In these cases, it’s best to do some planning in the design phase of your experiment ...effects with Sanger sequencing, provided you have some idea of where in the genome you might expect off...

...replication. However, this approach could permit some viral protein production, so researchers are also...Cas9 cut site, leading Wang et al. to conclude that some Cas9-derived indels may not ablate viral function...would open up many new therapeutic possibilities for some of the world’s most common diseases. References...

...different naturally occurring strains of E. coli, some of which are deadly to humans. The majority of all...for growth rpsL Mutation in subunit S12 of 30S ribosome Confers resistance to streptomycin Tn10   Confers...Additionally, Table 2 provides a quick reference for some of the popular strains, their genotypes, and their...

... Drosophila or development studies, you may hear some transcription factor and promoter pairs referred...behave very similarly?” The simple answer is that sometimes it is necessary or desirable to control or look...Conclusion This post contains only a brief overview of some of the most common repressible promoter systems,...

...enables the design of primer pairs that PCR amplify in some states but not others. Because samples of many registers...output plasmid (single copy, bacterial artificial chromosome backbone) contains the register. Depending on... that, depending on the context and application, some inducible recombinase systems may work faster or...

...offspring. Although powerful, this technology is cumbersome to use, not that efficient, often takes more ...must beware that not all gRNAs are created equal. Some work better than others. There are a variety of ...other gRNA design tools available and you can get some additional advice from John Doench’s gRNA design...

...select, and isolate clones. This procedure is cumbersome and inefficient, as it relies on random integration...there is no selection, you will need to screen somewhere between one hundred and one thousand clones to...boost efficiency over transfecting with plasmids in some cell lines, most likely due to DNA toxicity. More...