...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 ...

...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...

...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...

...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...

...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...

...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...

...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...

...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...

... 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,...

... themselves from one location and then insert somewhere else (Fig. 2). Class II TEs have LTRs on both ...Applications of transposons Now that you’ve learned about some of the popular transposon systems, let’s take a ...and cell therapy, and have potential to overcome some of the limitations of viruses. TEs have a large ...

...containing gaps or DNA base pair mismatches (8-11). For some applications, there cannot be any end-joining activity...those containing one or more mismatches. However, some ligases can ligate mismatches to a significant degree...pyrimidine:purine mismatches. Helix stability also plays some role, and mismatches with more hydrogen bonds are...

...swarming around S. aureus and eating the bacteria. Some of the many questions scientists have been able ...used in your experiments, experience has shown that some are more reliable and more efficient at tracking... Tips on Choosing Your Fluorescent Protein Learn Some of the Pitfalls of Working With Fluorescent Proteins...

...therapies themselves can be made more effective. Some important factors researchers are beginning to focus...broadly activate the immune response against tumors. Some chemotherapeutic agents induce immunogenic tumor...deaths that help activate tumor responses. Similarly, some radiotherapies have caused increased death receptor...

...approaches are commonly used to produce KO cell lines, some of which are commercially available. Various proteomic...multiple techniques. For example, we have found that some antibodies perform well in a PFA-saponin fixation...). The approach maps onto the five pillars, with some specific recommendations for how to use these in...

...will deliver some fabulous photos again this year! Why #DecktheLab? Help us spread some cheer by snapping...

...on the basics of viral vectors, applications, and some new resources available from Addgene. There’s a ...vectors In addition to “going viral,” we also have some other exciting news. Last week, we launched our ...

...after all a place that encourages people to take some time out every once in a while – just read Melina... people in a better mood. Laughter itself can sometimes even count as demanding exercise, and, of course...

...available online if you prefer the DIY method. As some proteins aggregate at 95 °C, you may want to check...preserve sample, check the ingredients before using, as some contain Tween-20, which can strip proteins from ...non-specific binding; lot-to-lot variability Some recombinant antibodies Not always available; not...

...received a festive makeover today✂️❄️🦠thanks to the awesome templates by @CVRHutchinson @CVRinfo. Check them... to everyone who decked their lab and brought us some holiday cheer!   ...

...export license application for rabies materials! Some of the materials in our repository, including rabies...license for rabies materials may be required for some countries and may not be required for others. You...

... about their career paths, future goals, and get some useful career advice.  Our first video, featuring...and interesting content. Wish we had a video on something that you don’t see here? Give us suggestions in...

...3 choices), we'll be in touch with you soon with some brand-new Addgene swag!   Last month, we introduced...submission is included in the final four will receive some rockin’ newly designed Addgene swag. Good luck and...

... I went to college for microbiology and then did some independent research my junior year and discovered...other companies. It's one of those "every day is something different" type of jobs, which really suits my...

..., welcome to our new and improved site! Here are some of the exciting new features on the blog that will...will give you a better experience. Looking for something? Use the navigation menu. The new navigation menu...

...graduate student life. Senior graduate students can sometimes be elusive or have a certain mystique that makes...University of California, Los Angeles. She enjoys chromosome biology and reading Ben Vincent's tweets.    ...