...started learning proper cell culture technique, I tried to find an app that could help me with calculations...information in the app is restricted to the NEB inventory only, the restriction enzyme sequence resource...This post was contributed by guest blogger Sean D. Stacey. Opinions on the apps discussed in the article...comes in handy when I use protocols that require restriction enzymes or PCR. The app contains a double digest...digest tool that is only helpful for NEB restriction enzymes since the instructions use NEB buffers. Nonetheless...name to discover all the information about that restriction enzyme or share it via email with a colleague...colleague. The app is also great if you’re adding a restriction site to a primer, you only need to skim the alphabet...

...tools are abundant! In this post, we will review tried-and-true protein expression control tools as well...

...marker at non-physiological levels. Scientists have tried varied approaches to combat these issues. One method...maintained when grown on media containing the missing nutrient. Scientists can exploit these host mutations by...auxotrophy. When grown on media NOT containing the nutrient, the host cells will die unless they have incorporated...  Marcy Patrick...

...these elusive teachers, supporters and advisors? I tried to start a more formal mentoring program at my company...They all found the support of the group to be intrinsic to their success. Their stories made it clear ...Goal setting is easier with more perspectives contributing to the process Senior people (of which there...

...most widely known of these products is TRIzol® (also called TRI Reagent®, RNAzol®, QIAzol®  and sold by...homogenate and shaken. After a quick centrifugation (not ultracentrifugation!), the layers of phenol and chloroform...After homogenization in TRIzol®, insoluble material is removed via centrifugation and the supernatant is... loaded onto cesium-chloride gradients and ultracentrifuged for up to 20 hours to separate the intact ...have, access to one or more large, expensive ultracentrifuge.  Figure 1: An outline of the different...out to develop a protocol that skipped the ultracentrifugation altogether. Chomczynski and Sacchi showed... by many different companies). TRIzol® is an all-in-one acid-guanidinium-phenol solution that combines...

...writing three years ago on a field trip in Indonesia. Since then, I have tried many things: photography, making...This post was contributed by guest blogger Sarah Schmidt, a Marie Curie Fellow at The Sainsbury Laboratory...hosted by Nature), and Addgene are open to guest contributions. If you decide to start your own blog, check...

...confused, bored, or particularly excited about? If you tried tweaking your pitch throughout the conference, what...

...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...

...AiE0621h SYFP2 Olfactory tubercle and Fundus of striatum Striatum 214849 pAAV-AiE0621h-minBG-iCre(R297T)-BGHpA...AiE0621h Cre Olfactory tubercle and Fundus of striatum Striatum 191717 pAAV-3xSP10ins-AiE0367h_C2-minRho-SYFP2...using various delivery methods including intracerebroventricular (ICV) delivery in newborns, and retro-orbital...Isocortex Hippocampal formation Cortical subplate Striatum Pallidum Hypothalamus Cerebellum Whole brain Addgene...AiP13038 AiE0743m_3xC2 SYFP2 Cholinergic interneurons Striatum 214848 pAAV-AiE0743m_3xC2-minBG-iCre(R297T)-BGHpA...AiP13780 AiE0743m_3xC2 Cre Cholinergic interneurons Striatum 214858 pAAV-AiE0873m_3xC2-minBG-SYFP2-P2A-3XFLAG...AiP14496 AiE0873m_3xC2 SYFP2 Cholinergic interneurons Striatum 214868 pAAV-AiE0873m_3xC2-minBG-iCre(R297T)-BGHpA...

...BGHpA (Alias: CN5140) Expression of CoChR-EGFP in striatal cholinergic neurons Jonathan Ting Viviana Gradinaru...BGHpA (Alias: CN4825) Expression of iCre(R297T) in striatal medium spiny neurons Jonathan Ting Viviana Gradinaru...protein) plus separate nuclear FLAG epitope in striatal cholinergic neurons Jonathan Ting Viviana Gradinaru...BGHpA (Alias: CN4361) Expression of jGCaMP8s in striatal indirect pathway medium spiny neurons (D2 MSNs...BGHpA (Alias: CN4360) Expression of jGCaMP8s in striatal direct pathway medium spiny neurons (D1 MSNs) ...BGHpA (Alias: CN4035) Expression of CoChR-EGFP in striatal indirect pathway medium spiny neurons (D2 MSNs...BGHpA (Alias: CN4033) Expression of CoChR-EGFP in striatal direct pathway medium spiny neurons (D1 MSNs) ...

...biosensor for cyclic-di-GMP measurement Asymmetrical distribution of the second messenger c-di-GMP upon ...redox-sensitive ratiometric sensor roGFP for mitochondria or cytosol Hypoxia triggers subcellular compartmental...depdent on Cre, Flp, or Vcre Comprehensive Dual- and Triple-Feature Intersectional Single-Vector Delivery of...imaging (GECOs), including blue, green, red, and ratiometric indicators An Expanded Palette of Genetically...10.1117/1.NPh.11.2.024207. Robert Campbell Calcium Ratiometric calcium biosensors from nested reporters of green... green- and orange-emitting proteins Ratiometric Matryoshka biosensors from a nested cassette of green...anisotropy Cameleon calcium sensors Single-color, ratiometric biosensors for detecting signaling activities...

...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...

... 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 ...

... 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...

...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...