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...extension can usually ensure that the 3’-base(s) do not form secondary structures. The introduction (or ablation...plasmid (usually from incomplete DpnI digest) can also form colonies under these conditions. Screening If a ...constructs), in functional analysis of proteins (to perform alanine scanning mutagenesis or targeted substitution...methylation-dependent endonuclease (i.e. DpnI), and bacteria are transformed with the nuclease-resistant nicked plasmid (the...the adenosine of the GATC recognition site. Transformation: After the PCR reaction, no ligation is required...required since the E. coli you transform your PCR products into will efficiently patch up the DNA. The ...parental plasmid provides a mean for selecting for transformants which have taken up the mutagenized plasmid....

...IT6, and IT10 expressed well in cells and did not form undesirable aggregates. Using budding yeast as a...HA, and Myc can adopt a number of different conformations and thus potentially interact with and alter...compromise protein folding and function. Using bioinformatic analyses, they also selected for other key properties...

...Morphology is a simple and fast to evaluate. iPSCs should form compact colonies that are comprised of cells with...functional tests. Cells can be differentiated in vitro to form embryoid bodies which are compact balls of loosely...offspring demonstrates the ability of the iPSCs to form functional germ cells. The highest stringency test...after differentiation has been linked to tumor formation in iPSC-derived and iPSC-transplanted mice. Cre-deletable...to purify the reprogramming proteins needed to perform these experiments. Assays for iPSCs Characterization...teratomas is the gold standard instead. Teratoma formation requires injecting iPSCs into immunodeficient ...

...there are multiple amplicons, or primer dimers forming, the curve will show multiple peaks.    (a...Addgene frown upon statistical laxness in all its forms. Reporting your data  Once you’ve done the work ...share it so others can provide you with the highest form of flattery - imitation! Use the MIQE guidelines...most machines have a protocol that automatically performs the normalization step.  Machines Most general-use...machine. (Teo, et al, 2002; see paper for more information on optimizing for LightCycler machines.) Multiplexing...ensure you’re recording and reporting the right information so others can reproduce your efforts. And if ... J; Wittwer CT. The MIQE guidelines: minimum information for publication of quantitative real-time PCR...

...website also provides a variety of short- and long-form written content about timely scientific topics, ...biomedical sciences and decided to offer a varied format event to keep the audience engaged. The day kicked... let us know in the comments section! More information about SITN: DayCon event website (includes archived...

...can have successful careers doing science in some form? There is a lot of talk about the state of scientist...still university research departments that have no formal career resources for science trainees at all. For...

...under blue light CRY2-CIB1 heterodimerizes or forms clusters under blue light Check out our Optogenetics ... Rab11, and Rab35 to understand their roles in forming the cilia that help establish left-right organization...heterodimerizes under blue light BcLOV4 undergoes a conformational change and binds the plasma membrane under ...leading to behaviors like lamellipodia or filopodia formation, cell contraction, or other morphological changes...publications, they described tools to control formation of lamellipodia and contractile stress fibers,..., Rab11, and Rab35 coordinate lumen and cilia formation during zebrafish left-right organizer development...

...for someone with HR expertise to join their newly formed compensation committee. The compensation committee...

... a variety of mammalian cell lines. Alternative forms of this vector contain GFP-tagged and un-tagged ...confusing to the uninitiated, contains much of the information needed to understand how to use it. TLDR: This... Zhang Lab's Nature Protocols Paper for more information). V2.0 This designation separates pSpCas9(BB)...

...actionability on the Addgene blog and how SITN short-form blog posts help readers think through current events...a source of scientific information for the general public as well as a platform for scientists to be viewed... for listeners to understand the dynamics of information flow. Some of the topics they've recently covered...Mani is a graduate student at Harvard University, former Co-director of Science in the News, and member ... received her PhD from Harvard University, is a former Co-director of Science in the News, and is a member...

...et al., 2016). It’s generally accepted that some form of homologous recombination (HR) occurs via this...genomes encoding part A and part B of the transgene form a heterodimer, then splicing of the pre-mRNA of ...vector had an improvement in muscle physiological performance.  2. Trans-splicing Overview: For trans-plicing...et al., 2005). Trans-splicing does not always outperform single oversized AAVs (Hirsch et al, 2013). ...

...production.   MANOVA The MANOVA is a multivariate form of ANOVA. Isn’t that what a two-way ANOVA test is...distribution. If your data is normally distributed, it will form (more or less) a straight line. If your data does... enough information. A post hoc test, done after your initial analysis can provide information on specifically...ANCOVA analysis. If you have performed a MANOVA/MANCOVA and want this information, multiple ANOVAs/ANCOVASs...play by the rules” and which we have certain information about. The assumptions and requirements that ...than one another, a two-tailed test should be performed. This test will assess if the mean of an experimental...tests, referred to as post-hoc analysis, must be performed. These will be discussed later. One and two-way...

... However, if repair material is provided (in the form of an oligonucleotide or plasmid), precise changes...still have much to learn from biology in all its forms! Many thanks to our guest bloggers, Wenning Qin...concept, straightforward to use, and robust in performance. In its natural setting, CRISPR-Cas9 is an acquired...

...exception. For the past 10+ years, Addgenies have formed teams, chosen a theme for the costume, and did ... the snail, the larvae reach the eyestalk and transform them into a pulsating display imitating a moving...lethal, associated with defective exoskeleton formations. These genes were appropriately named as spook...grasshoppers and crickets. When it’s ready to transform into an aquatic adult, it forces its insect host...

...base pairs to the repeat section of the crRNA, forming the repeat:antirepeat duplex. tracrRNAs also contain...tracrRNA sequences in a “single-guide RNA” that performs both functions. Cas9 cuts both the target and ...

...While in Grad School How to Lead a Great Meeting Form Your Own Peer Mentoring Group...interdisciplinary seminars, such as those in bioinformatics or chemistry, even if your primary focus lies...

...engineered gene clusters that encode proteins that form bacterial gas vesicles. These ~200nm long vesicles...by an inexpensive and widely available imaging platform: ultrasound. Challenges of in vivo imaging with...ultrasound imaging. Ultrasound is an attractive imaging platform for in vivo imaging experiments because  ultrasound...Following in vitro characterization, Bourdeau et al performed a proof-of-concept experiment to demonstrate the...

...guidance. Every plasmid shared, every collaboration formed, and every breakthrough enabled brings us closer...data for over 150,000 plasmids, empowering bioinformaticians, data scientists, and developers to unlock...Schofield, C., … Laflamme, C. (2024). A consensus platform for antibody characterization. Nature Protocols...

...Overexpressed proteins also have the potential to form aggregates and may be toxic to the cell. Here comes...What can you do with SunTag? Tanenbaum et al. performed a variety of different experiments in their paper...

... collection by filling out our Suggest a Plasmid Form. We look forward to helping microbiologists from...sharing and was also a great place to catch up with former colleagues! I ran into past ASM President Tim Donohue...