...shipment processes. After reading a publication describing how short term storage on dry ice can shift sample...

...on social media or the phones hiding under their desks than listening to me. When I got home, I saw my ...

...next-generation sequencing (NGS). For gRNA libraries, you’ll design primers to create 200-300 bp sequencing products...

...bacterial proteins that bind serotonin. Unger redesigned the binding pocket of the periplasmic binding...

...producer cells and purification protocols often describe a process that begins with cell lysis to release...

...ability to influence editing efficiency. In 2022, despite the increasing effectiveness of CRISPR/Cas9, scientists...

...characterization of the new variants is available here. Upgrades to iGluSnFR3 To improve upon the SF-iGluSnFR, ...

...store your plasmid DNA Learn about the different grades of plasmid DNA preps Resources on Addgene.org ...

...on Twitter where each poster presenter had four “slides” to use for their poster, eliminating the need ...

...to note that it’s the depositing institute that decides the terms of distribution for their plasmids. ...

...retroviral CRISPR knockout library using gRNAs designed by the Kosuke Yusa Lab. This retroviral library...

...with NEB enzymes, the app is quite helpful and provides a list of directions including which buffers to...

...of R bodies R bodies have been studied for many decades, and they were known to be resistant to detergents...

...Define the scope of the meeting: Are you meeting to design an entire experiment and delegate tasks to lab ...

...University of British Columbia, Student Services - includes videos on general resume writing, as well as additional...

...run on an agarose gel. The resulting gel image includes a 1kb ladder (lane 1) that has bands ranging from...

... a Good Mentor for Scientists". This post also includes a useful list of questions for a candidate to ...

...even if science writer Ed Yong’s book “I Contain Multitudes” has me considering the microbiome more often...

...different phenotypes. For example, this plasmid encodes a green fluorescent protein gene which makes the...

...2A-Puro (PX459) - This mammalian expression vector encodes both SpCas9 and your gRNA. It’s the former version...

...Bhopal. All illustrations / figures featured were designed by our team member Kallol Mozumdar. Sreepadmanabh...

...about a specific virus? We have you covered with guides on lentivirus, retrovirus, adenovirus, and AAV....

...focusing on molecular biology and metabolism. He now guides fellow PhDs in their careers with actionable steps...

... that you know how to read flow plots and have designed your first flow panel, you’ll load your samples...

... another way, only cells or organisms with the desired phenotype will survive and be assayed at the end...

... don’t overdo it (some of my best studies were designed over late night poker games at Gordon Conferences...

...have been a model organism for plant research for decades, and the biochemical pathways involved in central...

...There are four biosafety levels. This protocol provides information for both biosafety level 1 (BSL-1)...level has different safety requirements. BSL-1 is designated for those working with microbes that don’t cause...Staphylococcus aureus or Vibrio cholerae . BSL-2 includes all of the precautions needed in BSL-1, along ...before and after working in the lab. Ensure that a designated chemical waste accumulation site is present, ...(MSDS). While You Work Dispose of sharps in a designated sharps container. Sharps include anything that... drain. Solid BSL-2 waste can be collected in designated biohazardous waste containers that can be autoclaved...important! Disclaimer: The SOP presented here has been designed by the Addgene nonprofit plasmid repository and...

... Procedure Design your plasmid and order primers (see figure to the right). When designing your plasmid...and enrich for correctly assembled plasmids by designing primers to split an antibiotic resistance gene...and yield. If there are significant amounts of undesired product, gel-purify DNA segments. Otherwise, PCR... High-Fidelity DNA Polymerase - incorporates nucleotides to “fill in” the gaps in the annealed DNA fragments... (<150 bp). Pro-Tip Please note that the way to design the “stitching” primers and the amounts of primers...window) . NEB has other resources, such as a primer design tool (Link opens in a new window) . Gibson Assembly...

...created. Additional design rules can be found at the Broad Institute's TRC shRNA Design Process . To minimize...Consortium A.2 Map of pLKO.1 A.3 Related plasmids B. Designing shRNA Oligos for pLKO.1 B.1 Determine the optimal...sites in place of the stuffer. The AgeI site is destroyed in most cases (depending on the target sequence....9kb stuffer, visit www.addgene.org/10878/ . Description Vector Element U6 Human U6 promoter drives RNA...the pLKO.1 TRC-cloning vector. Addgene Plasmid Description pLKO.1 – TRC control Negative control vector ...website and “search for “pLKO”“. Back to Top B. Designing shRNA Oligos for pLKO.1 B.1 Determining the Optimal...accessed after a free registration. Guidelines for designing siRNAs with effective gene silencing: Starting...

... phosphatase activity: In yeast, the PHO8 gene encodes a vacuolar alkaline phosphatase. Normally PHO8 ...

...the postdoctoral population which keeps growing despite a lack of available academic positions.   Many...

...novel RNA-protein interactions, Ramanathan et al. designed an RNA bait containing three well conserved but...

...these difficult pieces: “It’s probably the best.” Besides working through the tough assignments, the skills...

...to the development and spread of cancer cells. Besides these stromal cell modifications, stromal cells...

...affinity, many indirect antibody protocols are designed to take place over multiple days. Increased run...

...Allewell and Sarma, 1974). Additionally, ThermoFisher provides a protocol on how to integrate RNAlater® with ...

...talking about here? Many of the Fab fragments described above can be used for similar applications and...

...down your own results; are they the same as the description the authors give? Ask yourself if the results...

...Activation of membrane receptors and signaling cascades conceivably allows the genome to sense the impact...

...restriction enzyme digest (subcloning), including design and experimental procedures....easily move YGOI into a mammalian expression vector. Design (Choosing enzymes) Many DNA analysis tools, including..., but do not cut within your insert Are in the desired location in your recipient plasmid (usually in ...your recipient plasmid as well as a specifically designed test digest later to verify that the insert was... fear. You have other options, such as: Adding desired restriction sites to flank your insert : You can...restriction enzymes that cut within your insert. Adding desired restriction sites to your recipient plasmid : You... orientation of your insert, so you may want to design a diagnostic digest for this purpose. Congratulations...

...to a lay audience. My lay paragraph typically includes a couple of sentences of introduction to my area...

...guanidine moiety at the end of its side chain. Decades ago, other labs discovered that the in vitro functions...

...for faster detection of RCA. 2nd Gen AdVs were designed to have increased cloning capacity, decreased  ...

...specific protein-protein interactions, SplitBioID provides a candidate list of proteins that are part of ...

...Biology Tips from the Repository Trenches: Using Barcodes to Track Samples Resources at Addgene.org Find...

...ELISA is a modification of the sandwich ELISA as described here, the direct and indirect ELISA (more details...

... Parts. This library delivers 2 randomly paired guides per construct and allows a reduced scale in screens...

.../35++ Adenovirus Vector Expressing Anti-CRISPR Peptides Decreases CRISPR/Cas9 Toxicity in Human Hematopoietic...

... I can have on an organization beyond my job description.” Our resident bagel provider, source of energy...