...answer should come soon, but not instantly. Addgene tries to get back to customers within 24 hours, and we...time at Addgene, I’m confident that these tips and tricks will apply to any biology-related customer support...

...’t tell if the bags contained flora or fauna. I tried to find out what certain ingredients might be used...very welcoming team from China’s Ribobio, the illustrious speaker list attracted a host of sponsors based...and depositors. China is in one of the top 10 countries in number of plasmid requests from Addgene. ...

...answers come sooner rather than later. Once you’ve tried it, let us know how xCas9 is working for you in ...another method to create point mutations, also has a strict editing window about 15 nt upstream of the PAM....included continuous outflow of phage, increasing stringency and promoting survival of only the broadest-targeting...

...keywords close enough... If in the past you have tried to combine multiple terms in an Addgene plasmid ...plasmids from the James Trimmer Lab published in eLife. Addgene is currently distributing 94 plasmids expressing...Availability to Industry: Restrict search to plasmids which can be distributed to industry research scientists... GCaMP containing plasmids are available for distribution. If your experiment involves delivering a calcium...

... brain tumor biology. When not snipping DNA, he tries random things (101 lessons on Spanish, C++, guitar...DEMONSTRATE GENOME EDITING BY THIS TOOL This post was contributed by guest blogger Pooran Dewari. Any views in ...performs Sanger sequencing on select regions of all distributed plasmids as part of quality control, but does...with the NgAgo system indicates that it is very tricky to use and fussy to optimize. One reason could ...inconvenient. Another possible reason for the trickiness of NgAgo is the inability of mature NgAgo protein...enzyme." DNA-scissors (blog). Blog website. 4. Hsu, Patrick D., Eric S. Lander, and Feng Zhang. "Development...

... be a shortage of amusing reports where someone tries to “optimize” a particular standard lab protocol...technical knowledge and judgement. For particularly tricky cases, team discussions can often lead to otherwise...

...     Over the last decade, my peers and I have tried to move away from calling non-academic careers and...hands-on lab opportunities. You can get your hands in nitrile gloves again! The opportunities are there once ...

...populations can arise very quickly following the tried and true process of evolution. Watch this video ...This post was contributed by guest blogger, Jennifer Rivers, a research microbiologist specializing in...resistant to antibacterial drugs. Resistance can be intrinsic, meaning a basic characteristic that naturally...resistant somehow overtime. One fantastic example of intrinsic resistance would be Mycoplasma species’ resistance...possess cell walls. As a result, Mycoplasma are intrinsically resistant to this class of antibiotic. Acquired...be unharmed by the compound it produces (think intrinsic resistance), but surrounding microbes would be...resistance genes survive and multiply. Factors contributing to the development of resistance include (Figure...

... glycine pH 2.5 and immediately neutralized with Tris pH 8-8.5. Low pH glycine is very effective at disrupting...and blue triangles. 2) An antibody immobilized on a bead binds specifically to the blue triangle protein...not the other. 3) The beads are collected by centrifugation or a magnet. 4) Unbound proteins are washed...target protein. The beads can be collected by centrifugation (agarose beads) or a magnet (magnetic beads...particularly advantageous because they do not require centrifugation between washes which can cause loss of beads...containing non-ionic detergents, such as NP-40 or triton-X, to preserve native protein conformations and...proteins, including those in the nucleus, more stringent buffers may be needed. Since many factors, including...

...cells to let the virions out. We recommend using a Tris-based lysis buffer and sonication.  The obtained...density gradient centrifugation (e.g., iodixanol) using a high-speed ultracentrifuge. During the spin,..., and the two helper plasmids, commonly called “triple plasmid transfection” (Figure 1).    Figure 1:...ddPCRAAV Purification by Iodixanol Gradient UltracentrifugationAAV Production in HEK293 Cells...

...If you’ve heard of containerization, but haven’t tried it or are confused by the concept, don’t worry–you... developed for a singular purpose (ex. adapter trimming, quality score analysis, k-mer counting, or sequence...first n items. To be sure, we went through much trial and error to learn the scripting language, but our...process. For example, our bbduk process created trimmed FASTQ files as output, which we specified as input...

...game show “The $25,000 Pyramid” where one player tries to get the other to guess a category by listing ...

...was first described in 1966 by Austrian physician, Andreas Rett, who tried to raise awareness and published...binds to methylated CpG dinucleotides and CAC trinucleotides during neuronal maturation (Tillotson & Bird...in which too much of the MECP2 protein is as detrimental as too little, resulting in MECP2 duplication...advance their respective gene therapies to clinical trials in 2021. Figure 1: Schematic of the human...

...for most purposes, but what do you do when you’ve tried a common strain and don’t get the desired level ...inactivated native restriction/methylation system. This means the strain can neither restrict nor methylate...expression of T7 RNAP from the genome; does not restrict methylated DNA fhuA2 lacZ::T7 gene1 [lon] ompT...adding the inducer. Growth Media: Changing media is tricky, because there can be a trade-off between growth...about your favorite protein expression tips and tricks! Leave a comment below or browse Addgene’s collection...

...flow strips that contain streptavidin which can bind to the biotinylated targets. They also tried to confirm...lateral flow strips. Since the probes are tagged with streptavidin, the lateral flow strip first captures...This post was contributed by Shravanti Suresh from Iowa State University. Since its appearance, SARS-... this virus has become an utmost priority for countries around the world and to do so, the WHO recommends...testing, tracking, and social distancing. With countries like South Korea, one of the earlier epicenters...be captured by an antibody further along on the strip. Highlights: Of the 83 known COVID-positive samples...viruses. Unlike Cas12a, Cas13a does not require strict sequence preferences at the target site, while ...

...originally being developed for A → G edits, researchers tried inhibiting MPG/AAG to improve editing efficiency...

...get them below a given threshold even when you’ve tried everything listed here. For some applications, this...CRISPR to (hopefully) be used in human clinical trials. An off-target rate of 5% is far too high when ...

...’s catalog — from the recently developed to the tried-and-true classics. Check out our Biosensors Collection...biosensors new to Addgene this year. Feel the love with MTRIA sensors One of the latest additions to Addgene’s...TRansmembrane region-to-IntrAcellular loop linkers” (MTRIA for short), where a fluorescent protein is inserted... ligand (Ino et al. 2022). The oxytocin sensor MTRIAOT enabled real-time observations of oxytocin dynamics...earlier tools. They also produce more than 20 other MTRIA sensors based on GPCRs recognizing various neurotransmitters...ligands - almost A to Z! - from ATP to urotensin. The MTRIA sensors will be useful for investigating neuronal...sensors for these and other targets. Figure 1: MTRIA biosensors are generated by inserting a circularly-permuted...

...heard stories and read papers wherein a researcher tries 5 separate experiments at once to get confirmatory...procedures like pouring a plate or running a gel have intricacies that aren’t always mentioned in static written...

...Shortly after though, three groups independently tried reproducing these results and determined that even... expression. References: Allocca M, Doria M, Petrillo M, Colella P, Garcia-Hoyos M, Gibbs D, Kim SR,...