...consideration is: The number of inserts ...as some methods are more amenable to combining multiple DNA fragments... a high-level comparison of popular cloning methodologies, with a special emphasis on these five categories...Please note that many of the following cloning methodologies have been extensively described in other articles..., Golden Gate cloning might be for you. This methodology relies on versatile Type IIS restriction enzymes...option to consider when time is of the essence. This method requires a vector backbone containing a topoisomerase...choice, because the T4 polymerase needed for this method is relatively inexpensive and can be obtained for...approach, you might want to give Gateway a try. This method, developed by Invitrogen and based on lambda phage...

...cause demethylation?  Dividing cells depend on DNA methyltransferase 1 (DNMT1) to copy methylation marks...slightly smaller population, something else was also suddenly cool: DNA methylation. In 1983, Andrew Feinberg... proteins are not demethylases; they are dioxygenases that oxidize 5-methylcytosine to create rare DNA...complete DNA demethylation can be achieved by dCas9 alone, without TET (or any demethylase) fusion. This...site. So, when DNMT1 methylates the genome during DNA replication, it doesn’t methylate sites blocked by ...the original methylated strand is diluted to undetectable amounts: ergo, DNA demethylation! With optimization...optimization, this method can produce nearly 100% demethylation without the confounds inherent to dCas9-TET...

...Nielsen lab’s CRMAGE system is a fast, multiplexable method that combines CRISPR and recombineering-based MAGE... template or non-template strand, among other parameters. .  Stanley Qi lab's CRISPRi plasmids Stanley...based CRISPRa In the search for a CRISPR activation method in E. coli, Jesse Zalatan’s lab used an approach...as a collection of organisms, their genes, and metabolic processes, occupying the gut. Many human diseases...bacterial strain introduced to the population as methods of introduction. Engineering the bee gut microbiome...

...fragments. Know your cloning methods There are two other common cloning methods that are easy to get mixed...assembly was one of the most commonly used assembly methods (Kahl 2013). However, Gibson assembly isn’t ideal...linkers) which brings modularity to overlap-directed methods (Casini 2014). In this case linker sequences are... seamless cloning into a vector where no other methods were available. A second example of using Gibson...Gibson assembly improves on two aspects of the SLIC methods. First, it uses a dedicated 5’ exonuclease instead...Gibson, there are yet more homology based assembly methods that have been described--CPEC (circular polymerase...) but actually work by significantly different methods. Golden Gate cloning does result in the seamless...

...viral protein EBNA-1. Methods for reprogramming hiPSCs from PBMCs While the method of choice for reprogramming...somatic cells, the common goal of all reprogramming methods is the expression of the reprogramming factors-...derivation of clinical grade hiPSCs, reprogramming methods that do not involve modification of the original... clinical field [4]. Three major reprogramming methods are considered when generating hiPSCs for clinical...reprogramming factors To date, the mRNA transfection method has not been used successfully to derive iPSCs ...PBMCs efficiently [5]. However, the Sendai virus method is expensive, especially if used to make clinical...Episomal reprogramming is an ideal and cost-effective method for creating clinical-grade, safe, nonviral and...

...within a few days after you’ve met. Always include a note about where you met IRL (in real life) and perhaps...profile.”  The latter is much less creepy. If we’ve met before, I’d be much more likely to say  “Hey, thanks...This is also a great way to keep track of how you met the connection, as these notes are saved in the messaging...

...that interested in meeting movie stars, but when I met the astronaut Jim Lovell I was speechless (the club...protein - 1 RNA system Drs. Dounda and Charpentier met at a 2011 conference and decided to collaborate on...

..., one of the professors I met told me, “you can’t be interested in something you don’t know about”. I’... lab at UC Berkeley. Most of the scientists I’ve met began doing research in their undergraduate years...about the best way to answer those questions. I’ve met graduate students who enjoy the thrill of acquiring...mentors Many of the grad students and postdocs I’ve met have done things that I never even imagined before...Professor, Assistant Professor, Associate Professor Sometimes teaches graduate and/or undergraduate classes ...your way. Think critically about whether it’s something you want to dedicate your time and energy to. ...cultivate stronger relationships with my labmates. Sometimes what I thought was a dumb and very basic question...

...the conference Follow up with the new people you met at the conference. If you promised to send a paper...and ask for their input. Connect with people you met on LinkedIn, especially if they are not in academia... you are on a deep level. Right away you have something in common. Namely, the scientific question you...

...experimented with a group mentoring format where 1 mentor met with a group of mentees to get more “bang for the...Daniell describes a group of 7 women scientists who met every other Thursday for 25 years. They helped one...organizations as it brings a diversity of perspective. Sometimes it is helpful to find peers that share a similar...

...If you haven’t already, connect with those you’ve met at the conference. Networking All of the things we've... the conference Follow up with those that you’ve met. Connect with people on LinkedIn or Twitter, for ...day. After the conference If there’s a cool new method that you learned about at the talk, bring it to...up to my poster, I usually greet them and say something like “Let me know if you have questions or if ...’re familiar with the policies before posting something on social media. If someone is presenting unpublished...also helpful to write your Twitter handle on your nametag for the tweep up (and for the entire conference...conference moments on social media and if there was something you missed during the conference, you can try ...

...its position (number) relative to the start Methionine (Met) followed by the one letter abbreviation of...

...know anything about these people at the time, but I met some very accomplished scientists and some rising... connected people who otherwise might never have met. Binningsingletons.com will be the hub for all our...to someone new and ask for help. That was NOT something I would have done at my first six ASM meetings...provided moral support was invaluable.  Mixing metaphors and scientists - Culturing a scientific network... and Singletons can accelerate the networking timetable for the Singletons in their bins.  The next component...similar fields (ASM Microbe has eight tracks), something we didn’t have enough people to do in the pilot...

...this to be like any other research project. You’ve met someone whose career interests you or you want to...

...Zheng, a UX designer that she knew. “We had actually met at a lindy hop swing dance event,” Zheng says (networking...infect only one or a few species of bacteria and sometimes, only a subset of strains of the same bacterial...

...have been given the opportunity by Addgene. I have met some of the most genuine people who have shared amazing...to. Brandon Solis, marketing intern. Something that I struggle with as I’ve been thinking about...

...to work! Set up similarly to the online class, we met as a group for sessions covering individual portions... new technique, then type up the materials and methods. Over time that portion of the paper will be generated...

...booming, but, over the years, many researchers we’ve met with have asked if we can help them share and distribute...

...will get the research funded,” she says. Baradaran met Addgene Executive Director Joanne Kamens several ...for Biomedical Research. Drug development was something Stegmeier has been interested in for a long time...

...need just from reading up on a topic, but I haven’t met any.” A few other tips: “Learn HTML, write a lot ...