...– to specific sequences of DNA has become commonplace. This ability has opened up a world of possibilities...the original paper says). In their hands, BsaI-HF has produced less reliable results. She also notes that...

...specific field(s). Most anyone who has spent any time in academia has likely had the opportunity to attend... one that no one sees a purpose in attending and has lost both its focus and critical mass of participants...their own knowledge of the field. Perhaps interest has simply waned. When attendance slumps below a certain...rebooted or disbanded all together once their purpose has been served. I hope that with some of the information...

...Biotech in May 2016 (1), NgAgo has received much attention and the plasmid has been requested by labs around...THE ORIGINAL NgAgo ARTICLE DISCUSSED IN THIS POST HAS BEEN RETRACTED AND FOLLOW UP STUDIES HAVE FAILED ...spotlight and take a moment to remember that NgAgo has only been available for genome editing for a few ... synthesis, which could be less desirable if one has to target multiple sites in a single experiment. ...

...popularity. A yellow flame indicates that a plasmid has been ordered more than 20 times; red, more than 50...available through Addgene. Publication: If the plasmid has been published, the paper in which it was first used... their new plasmid.  The Addgene Vector Database has more information about common vector backbones.  ...plasmid number here. Gene/insert: This describes what has been inserted into the backbone, and provides information...gene or DNA element of interest. If the depositor has indicated that there is a particular mutation found...

...limit what policy can do and, in this way, science has always been related to government and, consequently...face of evidence” (4). Frustration among scientists has been rising as evidence of climate change, the safety...unwanted results (14). Some loss of trust, I fear, has already come to pass. It may only be by advocating... As of April 13th the March for Science homepage has amassed nearly 500,000 ‘likes’, while their website...scientific funding (17). Federal funding for science has been decreasing for the last five years and the first...for Science: The website for the March for Science has the details for the march in Washington as well as...

...Counter-selection with sacB has revolutionized allelic exchange. Even now, only half our lab has switched over to..., 2014). In recent years, CRISPR-Cas9 technology has also aided in the creation of scarless mutations,...isolated are those where the allelic exchange vector has undergone single crossover and integrated into the...to pTOX. However, we feel it has clear advantages, and are hopeful that continued development of allelic...the lab of Matthew Waldor at Harvard. Now that he has an efficient allelic exchange system, he is so excited...

...more commonly known: the fruit fly. The fruit fly has been used in scientific research for over a century... a transgenic mouse line. Over the years the fly has become an ideal model organism to study an array ...development, genetics, and the nervous system. The fly has also emerged as an important human disease model ... described in 1993 by Norbert Perrimon’s lab and has been continuously improved upon since then. If you...Zebrafish (Danio rerio) Since the 1960s, the zebrafish has gradually become an important model organism. They...Caenorhabditis elegans) The worm, Caenorhabditis elegans, has been used extensively as a model organism since the...

...abundance. If a scientist wants to know if an organism has a particular gene mutation, southern blotting can... DNA is complementary to the probe (for example, has the sequence of the mutated gene the researcher is..., two DNA fragments are observed because the DNA has the site where MstII cuts. In a patient with sickle...DNA fragment is observed. Today Southern blotting has been largely replaced by real-time PCR to answer ... large amount of high-quality DNA, real-time PCR has several advantages including easier automation, higher...questions (see below). Importantly, although their role has changed, restriction enzymes are certainly not dead...

...refined to create Brainbow 3.2. Brainbow labeling has also been applied to cell marking and lineage tracing...example, Brainbow-1.0 expresses RFP when the construct has not undergone recombination. This default state was...neuronal pThy1-Brainbow constructs, Addgene also has two Brainbow adeno-associated viral vectors (AAV)...Drosophila and zebrafish. Brainbow in Drosophila has aided in the mapping of neural circuits, such as ...neuromuscular junction. In zebrafish, the method has become very useful in lineage tracing; “Zebrabow”...Lichtman estimate that Brainbow’s colorful labeling has decreased the mapping time for a given section of...

... first publication in 2012, the Nagai laboratory has assembled a collection of multicolored nano-lanterns... low luminescence By itself, Renilla luciferase has a low quantum yield, and when transiently expressed...using chemiluminescent proteins for imaging, but it has precluded widespread adoption of the technology. ...Nano-lanterns and your experiments The Nagai laboratory has a done a wonderful job characterizing Nano-lanterns...

...CRISPR/Cas9 technology has been a breakthrough for the genome-editing field. It has the possibility to treat... applications.” Li’s team noted that CRISPR/Cas9 has been used to generate mutations in a wide range of... expanded organoids. “CRISPR in adult stem cells has huge potential, because the genomes of adult stem...

...for the flow plots discussed in this post, voltage has been adjusted appropriately and compensation performed...baseline expression of a protein while the green (top) has an upregulated expression (shift to the right). The...overlaps with the baseline suggesting this sample has a comparable expression to the baseline/negative ...allow for better interpretation. Priyamvada Prathima has a research background in protein mutagenesis and ...

... safe to assume that anyone reading this article has a smartphone nearby. We tailor these devices to our...and a couple of other nice features. This app also has a great educational section. This section holds the...this MSDS app. The Chemical Safety Data Sheets app has a search function that allows the user to find an...memorized, but the structures are another story. AMINO has been very helpful when I’ve had to explain the properties...

...modified version of CAG promoter that the Zhang lab has tested in a variety of mammalian cell lines. Cas9...effective in certain cell lines. This point mutation has been corrected in V2.0. pSpCas9(BB)-2A-Puro (PX459...that, upstream of the Cbh promoter, this plasmid has a U6 promoter followed by BbsI restriction sites ...

...technology has been studied and used in assays for over half a century; the Addgene blog has an excellent...new aptamer also has a higher melting temperature than its Spinach ancestors, and has a lower magnesium...initially still unstable in eukaryotes, the Jaffrey lab has since developed new RNA-based scaffolds to contain...

... animal moving around to explore the environment has a different motivation (and brain areas associated... certain forms of drug addiction. When an animal has been exposed to morphine or other stimulating drugs...involved in locomotor sensitization? Previous research has shown that regions such as the nucleus accumbens ...dark compartment (Howerton at al. 2014). Addgene has several recombinase-expressing ready-made AAV vectors... tests.    Forced swim test The forced swim test has been used since the 1970s to test the efficacy of...

... in a town with a good public school system that has made strong efforts in the last 5 years to incorporate...curriculum. Our Parent Teacher Association (PTA) has been instrumental in supplementing that curriculum...some quick tips to get things going: If your PTA has a general meeting and a means to request funding ...without them! If you live in a school district that has a lot of parents in STEM careers, chances are they...nearly every talk I’ve given in the past 20 years has been aimed at grad students, postdocs, and other ...

...While much of CRISPR research has focused on genome editing, numerous discoveries have been made using...cleave the DNA. Much of the research using dCas9 has focused on transcriptional activation using a fusion...CRISPR-mediated purification of a specific genomic region has also recently been adapted to isolate genomic loci...tag systems including Protein A, 1xFLAG, 2xAM, and HA have been reported, as well as affinity purification...for many other CRISPR techniques, this method also has great potential for improvements and adaptations ...

...altering the sequence of the protein. As DNA synthesis has become quite cost-effective it seems easy for scientists...optimize your gene of interest? Codon optimization has become extremely useful in expressing functional ...neurons in vivo (Slimko and Lester, 2003).  Addgene has a large selection of codon optimized genes, so check...which have even been linked to some diseases. It has been reported that 5-10% of human genes contain a...//doi.org/10.1093/nar/15.3.1281 Slimko EM, Lester HA (2003) Codon optimization of Caenorhabditis elegans...

...decades, recombinant Adeno-associated virus (AAV) has been widely used to deliver genetic tools to the ...The most famous ones are influenza hemagglutinin (HA), myelocytomatosis viral oncogene (myc), simian virus...limitations of epitope tags and fluorescent proteins, it has been crucial to develop new tracers that enable accurate...are known to be notoriously difficult to label. It has also been shown that dendrites in the stratum oriens... provided better labels than fusions to 3 or more HA tags. 4. High-resolutions microscopy As indicated...