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...without exclusivity. Chapter 11: The Stem Cell Bank (CReM Bank) at the Center for Regenerative Medicine and...did I start doing this in the first place?" Many thanks to our guest, Darrell Kotton. Dr. Kotton is the...Boston University and Boston Medical Center. Special thanks to our sound engineer/producer, Eddy Page! Edward...

... 17] should cover it. Addgene: OK, thanks for your time, and thanks for depositing your plasmids with ...Howarth: Just the usual blood, sweat and tears. (Thanks to our healthy blood donor for keeping going while...be of use to others to Addgene. I am a big fan; thanks for the T-shirt! References: SpyLigase peptide-peptide...

...package. In a transfer plasmid, the transgene is flanked by two ITRs, and the ITRs are retained with the...Figure 1: Recombinant AAV vector with ITRs that flank the gene cassette, which contains a promoter, the...ITRs are symmetrical 145-nucleotide sequences that flank the ends of the single stranded DNA genome of AAV...in most plasmids. In an AAV vector plasmid, they flank the gene of interest. A single ITR consists of palindromic...

...results: On this page you can download an annotated GenBank File or a SnapGene file for the sequences listed.... If the depositor provided a custom annotated GenBank file, this can be found on the main plasmid page...relevant), and types. Types listed are standard GenBank feature types. Clicking on a particular feature...your next experiment. We would like to extend our thanks to you (our community of scientists) for suggesting...

...to have 50–80 bp of homology arms on each side flanking the change you'd like to introduce. We use Ultramer...plasmid donor, with two homology arms on each side flanking your desired insertion or mutation. Each arm is...pX330/PX335 system. Additionally, I would like to thank Le Cong for all his hard work creating amazing resources...community and sharing these resources with Addgene. Thank you, Le! This blog post was updated to include all...

...Harvard Medical School. We want to give a special thanks to the early-adopter scientists, who trusted Addgene...to hear the stories others have to tell.  "Also thanks for being Addgene. You guys rock." — Andrew York...Calico Life Sciences. Most of all, we’re incredibly thankful for all the scientists who have joined our community...your work, your research, and your discoveries. Thank you. You guys rock. ...

...didn't have the space to give proper credit to. Thankfully, as Addgene's resident blog editor, I could take...can refold correctly after denaturing.  Designed ankyrin repeat proteins, or DARPins, are genetically engineered...derived from a class of binding proteins known as ankyrin repeat proteins. DARPins typically consist of 3...an excellent option for many applications, and frankly, if ain't broke, don't fix it!  That being said...

...reproducibility and shared similar desires to take action. Thanks to Teytelman’s vast network and ability to identify...thinking about ways to address irreproducibility and thanks to Teytelman’s connection with Joanne Kamens, Addgene... them to connect? Through networking of course! Thanks to individuals in the group who were well connected...collaborate with this incredibly dedicated team. Many thanks to Tyler Ford, Caroline LaManna, Joanne Kamens,...

...the design of flanking overhangs. Although there are theoretically 256 distinct flanking sequences, sequences...their recognition sequence, creating four base flanking overhangs. Since these overhangs are not part ...can engineer unique enzyme recognition sites that flank their DNA fragment in an inverse orientation. This...

...recombination events, and that the orientation of the flanking loxP sites directs the Cre recombinase to invert...while FLP binds FRT sites. When a DNA sequence is flanked by target sites (floxed) in opposing orientations...Figure 1, top). The entire DNA cassette would be flanked by two pairs of target sites- one wild-type pair...placement of the two pairs of target sites. By flanking the reporter with one loxP site and one lox511...

...vector contains a neomycin resistance gene (NeoR) flanked by homology arms. The negative selection marker...” the resistance gene through the insertion of flanking LoxP sequences, you can remove the gene through...targeting vector such that a set of three LoxP sites flank the resistance gene and the targeted exon in the...has simply been replaced with the same sequence flanked by LoxP sites while the resistance cassette has...

...he describes the now infamous tale of Rosalind Franklin and her essential-but-unlauded role in the identification...within this section. The GFP story has its own Franklin in Doug Prasher, who originally isolated the GFP...science produced by the quieter, more reserved Franklins and Prashers will find it increasingly difficult...your career.  Acknowledgements: I would like to thank fellow Addgenies Joanne Kamens, Marcy Patrick, Aliyah...

...two sets of sgRNAs; A, the top 3 ranked sgRNAs, and B, the next 3 ranked sgRNAs. Dolcetto was able to discriminate... lab selected sgRNAs in this optimal window and ranked them based on their optimized sgRNA rules and potential... and Dolcetto library, sgRNAs were selected and ranked based on the previously established sgRNA rules...

...requested through Addgene. Their plasmid maps and GenBank files can be found on their plasmid pages. All ...terminators, promoters, and genes) were built with flanking primer binding sites (designed to have 20-30 nt...parts are also annotated on the plasmid maps and GenBank files uploaded to Addgene. We also recommend inserting...characterization of the recombinases at our disposal.  Many thanks to our guest blogger Nathaniel Roquet! Nathaniel...

...Hershey-Chase experiment. In the same year, Rosalind Franklin and her student Raymond Gosling took the first...diffracted image of the DNA. Finally, in 1953, Franklin along with James Watson and Francis Crick resolved...Inferno (2013) and the movie version (2016) stars Tom Hanks as the protagonist (Langdon). The plot involves ...?Download high resolution versions here.  Many thanks to our guest blogger Kartik Lakshmi Rallapalli ...

...primers that amplify the DNA sequence of interest and flank it with 50 base pairs of homology to the targeted...interest and 50 nucleotides of homology to regions flanking the target site).  The best applications for dsDNA...limited circumstances. An alternative approach is to flank the desired change with 4-5 silent changes in the...recombineering system? Share your tips below! Many thanks to our guest blogger, Beth Kenkel Beth Kenkel is...

...cable ties) received a festive makeover today✂️❄️🦠thanks to the awesome templates by @CVRHutchinson @CVRinfo...uDEE9PsspD#deckthelab #virussnowflakes #sciart #thankyouscience #scicomm pic.twitter.com/DWKWaPs0iY— Xi Chen...  Congratulations to all our winners, and a huge thank you to everyone who decked their lab and brought...

...importance of sharing information/knowledge Outro Many thanks to our guests, Melina Fan, Kenneth Fan, and Benjie...Officer. Prior to Addgene, Kenneth was an investment banking analyst at Credit Suisse’s Technology Group in ...Engineering from MIT, both in Computer Science.   Special thanks to our sound engineer/producer, Eddy Page! Edward...

...Addgene to succeed and to improve, so thank you. 500,000x Thank You! Have a question? Email: [email protected] our favorite people – you, our scientists!  Thank you for being part of our growing community! Our...

...accessible for scientists as possible.  He also thanked our many business partners, without whom Addgene...essential advice and service. Joanne shared a moving thank you email from a requestor in Iraq (Addgene's first...family, too. Here’s to another great 10 years!    Thank you to our guest blogger! Blugene, Addgene's friendly...