... and dsDNA. This is not true for all amplicons; if your amplicon is AT-rich or has a secondary structure...Allelic specificity Can be multiplexed Single amplicon validation technique Hydrolysis Probes (Taqman...Bustin, 2017 for a more in-depth review.)  Your amplicon should be between 70-200 bp long With ΔΔCT analysis... intensity should smoothly increase as the DNA amplicon transitions from single-stranded to double-stranded...final temperature increase. If there is a single amplicon, there should only be one peak, as everything ... the same temperatures. If there are multiple amplicons, or primer dimers forming, the curve will show...curves showsing (a) a single peak, indicating one amplicon and no primer dimers. (b) A shoulder peak indicating...

...problem was the creation of an Addgene lexicon. The base lexicon was and continues to be a list of commonly...every new Addgene employee gets “Lexicon Training.” Since the lexicon is a “living” document, I update...format” all received formal definitions in our lexicon. We created documents and presentations, gave real...technology in a consistent way. Just creating a lexicon was not enough. We also had to enforce and maintain...

...It is a well known fact if a multicopy variant of a gene exists, there must be assays that researchers...fluorescent probe and quencher to quantify specific amplicons.  Sybr has high background noise, due to the large... affected by AT-rich DNA and the length of the amplicon and it has a higher rate of false positives (Colborn...is not affected by AT content or length of the amplicon. As the probes can have several different markers...DNA quality and primer design, particularly for amplicons under 100 bp (Ma, 2015.) A schematic representation...confounding by AT-rich DNA and by variation in amplicon length. Am J Trop Med Hyg. 2008 Dec;79(6):887-...

... and W are known to have replicons with broad host ranges. Popular replicons from these groups include...Building broad host range recombinant vectors Replicons, which consist of a plasmid’s ori and all of its...2011 and Jain and Srivastava 2013. Family Replicon Compatible with Shown to work in IncN ...amylovora, E. carotovora subsp. Carotovora, E. herbicola, E. rubrifaciens, E. stewartii, Escherichia coli...In addition to incorporating broad host range replicons, recombinant vectors can be modified further to...

... any plasmid: the origin of replication/replicon. The replicon is comprised of the origin of replication...itself as it must to survive within cells. The replicons of plasmids are generally different from the those...regulation and can be manipulated with mutations in the replicon. For example, the pMB1 ori maintains about 20 ...amplifying more copies with the addition of chloramphenicol – make sure your growth conditions aren't working...

...apply a variety of CRISPR tools to dicotyledonous plants. The pDGE Dicot Genome Editing Kit contains Cas9... PubMed PMID: 25730490 New kit for editing dicotyledonous plants The Stuttmann lab generated the vectors...CRISPR knockout pooled library Parasites from the apicomplexan phylum cause severe human and livestock diseases...Despite their importance to global health, most apicomplexan genes remain uncharacterized. Recent studies...the contribution of each gene from the model apicomplexan organism Toxoplasma gondii to the infection ...uncharacterized genes that are conserved among apicomplexans and important for T. gondii fitness. Sixteen... cells, and one of them called “claudin-like apicomplexan microneme protein” (CLAMP) was shown to be important...

...General protein expression BL21 (DE3) pLysS* Chloramphenicol (pLysS) pLysS expresses T7 lysozyme to reduce...Expression of toxic proteins BL21 (DE3) pLysE* Chloramphenicol (pLysE) pLysE has higher T7 lysozyme expression...Expression of insoluble proteins  Rosetta2 (DE3)* Chloramphenicol (pRARE) Good for “universal” translation; contains...Expression of eukaryotic proteins Lemo21 (DE3)* Chloramphenicol (pLemo) Rhamnose-tunable T7 RNAP expression...T7 expression. The pLys plasmid contains a chloramphenicol resistance cassette for positive selection ...

...and drop an icon onto the workspace. The color and size of icons are adjustable. New icons can be requested...scientists. The web-based collection of over 20,000 icons is designed by professional medical illustrators...

...Nagai laboratory has assembled a collection of multicolored nano-lanterns for use in various applications...appropriate for use with Nano-lantern technology. Multicolor imaging using Nano-lanterns requires a linear...Expanded palette of Nano-lanterns for real-time multicolor luminescence imaging.” Proc Natl Acad Sci U S... of bright luminescent protein for real-time multicolour bioimaging.” Nat Commun. 7 (2016): 13718. PMID...

...What’s a science writer?” Learning about SciComm by doing SciComm Writing had always been a strength of ...right place at the right time when she started her scicomm career: “ I think living in Washington D.C. helped... scicomm...

... Cuneate tract, dCST= dorsal cortico spinal tract, vCST= ventral cortico spinal tract, dc= decussation... first segments of the spinal cord showing the cortico spinal tract, CST, spinal cord and cranial nerve...device (EMCCD) or complementary metal-oxide semiconductor (CMOS) sensors, it takes only a few milliseconds...

...flanking the editing site on either side of the amplicon. Based on the frequency of out-of-frame edits...deleted accordingly. Pro tip: make sure your PCR amplicon isn’t too large so that you can easily visualize...expect off target editing to occur. Luckily, in silico prediction analyses like CRISPRitz or CRISPOR can...CRISPRitz: rapid, high-throughput and variant-aware in silico off-target site identification for CRISPR genome...

... written jointly by Addgenies Brook Pyhtila and Nicole Waxmonsky Resource sharing shortens the time needed...This post was written jointly by Brook Pyhtila and Nicole Waxmonsky. References McDade J, Waxmonsky NC. Emerging... Nicole Waxmonsky...

...little more this October. Science favorites Musicophilia: Tales of Music and the Brain by Oliver Sacks...properties it has. As with other works of Dr. Sacks, Musicophilia includes case studies of relevant neurological...book! Far from a work of glib pop psychology, Musicophilia is accessible and fascinating, all without compromising...

...Massachusetts Medical School developed a dCas9-based multicolor labeling system to visualize chromosomal loci... U2OS cells.   Figure 2: CRISPR-Sirius tricolor labeling. (A) Diagram of sgRNA-Sirius-8XMS2, sgRNA-Sirius...plasmids at Addgene. References Ma, Hanhui et al. “Multicolor CRISPR labeling of chromosomal loci in human ...

...BioRender.com.   SLIC also opened up a new world of multicomponent assembly, as fragments with overlapping sequence...efficiency (~20%). Figure 2: Schematic of multicomponent assembly with SLIC. Multiple fragments are ...issue is sequence similarity. If fragments in a multicomponent assembly have 5′ or 3′ sequence homology to...

...cell have the same replicon and/or partitioning system. Let’s start with the replicon- the part of the ...The replication problem Plasmids with the same replicon are incompatible because they compete for the ...

...exchange vector. This vector contains sacB, an chloramphenicol resistance cassette, and the R6K origin of ...Valentovich 2014). We routinely clone the purified amplicons upstream and downstream of the desired deletion...products. To allow manipulation of naturally chloramphenicol-resistant strains, we have also included versions...

...These probes can be combined in two-color and multicolor NIR imaging.  Small miRFPnano A separate class...monomeric near-infrared fluorescent proteins for multicolor imaging across scales. Nat Commun 11, 239 (2020..., V.V. Near-infrared fluorescent proteins for multicolor in vivo imaging. Nat Methods 10, 751-754 (2013...

... better in one configuration than the other (Sandomenico et al., 2020). Figure 1: Comparison between...that help scFvs fold properly in the cytoplasm (Sandomenico et al., 2020). conversion to a cysteine free ...253 . https://doi.org/10.1006/jmbi.1997.1457  Sandomenico A, Sivaccumar JP, Ruvo M (2020) Evolution of ...