...tagging method that labels cells with heritable barcodes. Barcodes consist of an 8bp variable region and a 6...Direct reprogramming describes the process where differentiated cells are turned into a cell type of ...with the majority of cells failing to develop the desired identity. The development of single cell technologies...Morris Lab at Washington University in St. Louis provides a combinatorial indexing strategy that simultaneously...towards successful iEP reprogramming. MettI7a1 encodes an uncharacterized putative methyltransferase. ...

... been described in this blog. If your data does not meet the assumptions for the tests described above...experimental groups, but how do you determine if they deserve that oh-so coveted asterisk of significance? You...referred to as two-sample or independent, but these designations all refer to the same test. On some occasions...Correction and the Benjamini-Hochberg procedure are designed to reduce false positive rates and control for...error rate than the multiple comparison tests described above. Violated assumptions and statistics Statistical... making your statistical analysis plan when you design your experiment, instead of after you’ve already...

... lacks context unless it links to a verified description of that reagent online. Furthermore, if the link... link between the catalog number and reagent description is lost (e.g. a supplier is bought out by another...directly to the original article online. It is also designed to be persistent and exist online indefinitely...specific information about the plasmid: name, description, how the plasmid was generated, validated sequence...authors comply with journal policies, Addgene now provides an RRID and a resolvable compact identifier for...you directly to the lenti-Cas9-VQR plasmid page. Despite their differences, both of these PID formats for...

...or FP) is one of the most important steps in the design of an imaging experiment.  The choice is non-trivial...fusion protein, etc...).  There are many online guides – including an excellent series of posts by Joachim...information about fluorescent proteins.  It was designed with the goal of encouraging user contributions...while also maintaining a relatively strict database design (schema).  Registered users can submit new fluorescent...risk of inaccurate data submissions, so FPbase includes a versioning system that allows users to see when...for, the Table of Fluorescent Protein Properties provides a master list of all proteins in the database,...

...to a wide range of industries and is a highly desirable quality sought after by employers. These skills...correctly the first time. These skills are very desirable to employers because they save time and money ...Meteorologist Journal Editor Textile Technologist/Designer     Fieldwork and traveling Many people are...employers. Increasingly, computer skills are highly desired, as roles are becoming more reliant on technology...(Python, R, SAS, HTML, Matlab etc) are highly desirable in a range of roles, including: Science Reporter...find answers to include: Will you be sitting at a desk all day or be on your feet constantly in a lab? ...

...infectivity): The use of the VSV-G envelope provides the widest tropism or range of cells a virus can infect...The widespread use of γ-retroviruses (gamma-retroviruses) in cancer and stem cell research has prompted...lentiviral packaging methods. The three components described above (envelope, packaging, and transfer) are ...into the 293T packaging cell line. This system provides the greatest flexibility to pseudotype γ-retrovirus...nomenclature of the plasmid backbone is meant to be descriptive of the components found within the backbone. ...

...is not enough on its own to help you find your desired plasmid. In many cases, you will need a screening...you will need to sort through them to find the desired clone. Do your selection, then screen Why add a...what about identifying clones that contain your desired insert? This is where a  screen comes in. Let’s...blue-white screen, which relies on the lacZ gene. lacZ encodes the enzyme 𝛽-galactosidase which can hydrolyse...Sanger sequencing determines the precise order of nucleotides within a DNA molecule, in this case a plasmid... of the information you need, so you'll want to design a custom primer. Because Sanger sequencing can ...

...created SRIRACCHA in 2017 to enrich and identify desired CRISPR mutants (Wen et al., 2018). SRIRACCHA uses...constructs for the reporter to work and to make the desired genome edit:  A donor plasmid that contains a ...OH-T and doxycycline. A gRNA that targets your desired gene (whose sequence is also found in the reporter...well. The ratio of RFP to GFP expressing cells provides a real-time read out of Cas9 activity.  By using...integrated GFP reporter leaving you with just your desired genomic edit.  Find SRIRACCHA plasmids at Addgene...DSB is repaired by NHEJ which depending on the nucleotides inserted or deleted, can result in an in-frame...

...educational resources — including blog posts, protocols, guides, and collections — is a lot of material to sift... launched earlier in the year, as well as our redesigned homepage and our new search tool, are the results...a few of the strategies we used in our software design. In each case, we leveraged the team’s knowledge...per resource category to expose as filters and designed the search engine to return only the filters that... bits of information for a given resource, and designed the search engine to weight search results differently...'s new search Read to learn more about why we redesigned our homepage and mascot ...

... has precluded widespread adoption of the technology. In 2012, Saito et al. described the first Nano-...molecular and cell biologists over the last two decades, but their use can be limited by a few caveats....hours! The latest enhanced Nano-lanterns (eNL) described in Suzuki et al. improve the brightness of previous...substrate furimazine. In addition to the previously described applications, the enhanced signal of eNL allows...autofluorescence and a high signal-to-noise ratio. Despite the many benefits of Nano-lantern technology, there...

...assessed for the desired parameter. A droplet containing a cell that is positive for a desired parameter is...their observations. However, while flow cytometry provides analysis at the cellular level, it cannot be used...degree of scatter is captured by detectors and provides useful information about the cell. Light that ... traveling is called forward scatter (FSC) and provides information about the relative size of the cell...initially traveling is called side scatter (SSC) and provides information about the cell’s complexity. A cell...side scatter (SSC). Users can gate around their desired cell population for further analysis. In this example...

...into agriculture as species-specific pesticides and insecticides, an advancement over previous chemical...prone to degradation – those are the words that describe RNA! Double stranded? Well, that’s just for DNA...also known as RNAi, are short, dsRNA molecules designed to degrade mRNA transcripts. RNAi activates the...sequence is found (typically RNA that the RNAi is designed against), RISC can repress translation and/or ... and repetitive sequence. All of the R-loops described above play distinct roles in promoting cellular...

... could participate. Despite the group not having a public website, all the slides, handouts, and resources...science or even to discuss potential solutions. Despite all the talk, not much was being done to address...practices could be an actionable way to tackle widespread irreproducibility. Luckily, there were other ...thinking about reproducibility and shared similar desires to take action. Thanks to Teytelman’s vast network...practices could be an actionable way to address the widespread issue of irreproducibility. Use your network...

... molecule degraders” recruit the proteasome to destroy targeted proteins. We previously developed small...locus-specific knock-in of FKBP12F36V. Detailed protocols describing the use of these plasmids are available here....Nabet et al., Nature Chemical Biology additionally provides detailed information on cloning procedures used...degradation, and lead author on the recent paper describing the dTAG technology. Follow him on twitter @behnamnabet... 29581585. This reference contains the full description of the dTAG system. The dTAG system was employed...Conditional Mouse Models CRISPR 101: Cas9 Nickase Design and Homology Directed Repair Plasmids for endogenous...

...approaches to gender equality over the past several decades: “Fix the Numbers of Women” focuses on increasing...-art methods of sex and gender analysis and 2) provides case studies with concrete examples of how gender...using both sexes in basic research is excellent design), but they used all female stem cells—this was ...environmental factors. How to include sex in experimental design Recognizing the importance of analyzing sex, the... above, another excellent resource for research design is Kalpit Shah et al., “Do you know the sex of ...Corporations need to integrate these insights into product design. But eyes have been opened—and we cannot return...

...monitored over time by sequencing the barcodes in the population (all barcodes can be amplified using the same...with roughly 73 million semi-random, 30 bp DNA barcodes that are integrated into target cells' genomes...ClonTracer system, lentiviruses containing these barcodes infect the target cell population at low multiplicity...replicate populations, they found that similar barcodes consistently made up the majority of the resistant...resistant populations. In fact, 88% of the barcodes within the resistant populations were shared by at least...

...repository that will help epigenetic researchers custom design their own engineering plasmid tools in about a ...cloning, the Haynes lab’s Protocols.io protocol describes an elegant system that allows for the assembly... cells that make up the human body and G-baToN provides versibility in tracking and modulating the interactions... inhibitor response 1 (TIR1) expressed under a desired promoter with a specific spatiotemporal expression...interests via CRISPR techniques. This tool-kit includes an array of FPs as well as additional useful tags...libraries encode target sites flanked by randomized nucleotides in place of a protospacer adjacent motif (PAM...

...of 3’UTRs, the Arizona State University (ASU) Biodesign institute developed a 3’UTR database specifically...specifically for C. elegans. This database provides detailed information about C. elegans 3’UTRs structures, ...imaging processing. The combination of these methods provides unique opportunities to develop computational ...Addgene In addition to the wealth of open resources described above, there were many plasmid tools presented...self-excise, and covalently ligate their flanking peptides together (Wang et al., 2018).” With the split ...continuously curated as we continue to receive plasmids designed for the worm so check back frequently for what...

...degradation of the tagged protein. Degrons are destabilizing domains which will be identified by various ...proteasome for degradation. Like several other tags described here, the dTAG uses a small molecule, a tag, and...protein for imaging. Given its popularity and widespread usage, a proteolysis-targeting chimera (PROTAC... The auxin-inducible degron (AID) system is the oldest system reviewed here. The AID system relies on ...Thankfully, this is a hot topic with many resources! In Bondeson et al, they review all the tags discussed here...informed choice! References and Resources References Bondeson, D.P., Mullin-Bernstein, Z., Oliver, S. et al....

...amazing project. You’ve written your protocols, designed your experiments, and prepared your reagents. ...following cloning methodologies have been extensively described in other articles; so if you are interested in...homology. You might also consider FastCloning, first described by Chaokun Li and colleagues. Here you simply ...-generated insert. When mixed, topoisomerase I provides both the endonuclease and ligase function while...restriction enzymes. In Golden Gate cloning, the desired end construct does not contain the type IIS recognition...This feature biases the reaction in favor of the desired assembly to make the process close to 100% efficient...