...glycosyl groups, and other modifications to their bases in order to protect their DNA from host cell nucleases...proteins in the (all-too-common) event that homology-based searches return no candidates. Our strategy: Express...in the presence of certain modifications to DNA bases.  Ligate and transform your resultant plasmid libraries...Adapting drug approval pathways for bacteriophage-based therapeutics."Frontiers in Microbiology 7 (2016)...Raleigh. "Biosynthesis and Function of Modified Bases in Bacteria and Their Viruses." Chemical Reviews...

...plasmids and those without, we adjust our estimates based on the journal that published the articles.  That...Addgene plasmids as if they comprised 10 percent. Based on this new re-weighting, we can compare citations...Addgene is achieving its goal of making plasmid-based science more accessible…and accruing more citations..., we went to one of the largest bibliographic databases available, Web of Science. For each publication...number of citations the paper receives. From the database we pulled the journals from 2005 to 2015 in areas...

...you through both experimental and primer design based on fragment sequences and the polymerase used in...joining of multiple mutations spread over thousands of base pairs. Cloning sgRNAs Generating constructs to produce...Cloning Gibson Assembly and Other Long-Homology Based Cloning Methods Screen Plasmids by Colony PCR Addtional...

...plasmids used to generate these antibodies were based off of a large collection of mouse monoclonal antibodies...Antibodies?  One of the benefits of using a plasmid based system for expression of R-mAbs is that it can easily...Additional Resources on the Addgene blog:  Plasmid-Based Recombinant Antibodies  Plasmids 101: Secondary ...

...genomic SDM relatively straightforward, but plasmid-based SDM has lagged behind. While commercial kits are...PCR is used to splice together two PCR amplicons based on sequence homology, and the subsequent product...primers also have some homology to each other; ~17 bases is optimal. After being treated with DpnI to digest...

...limited to neuroscience. In fact, precise light-based control of biological processes is highly useful...better fine-tune the level of output gene expression based on the input light intensity. In addition to these...strains of E. coli, as well as with other plasmid-based systems, which will enable the assembly of larger...

...a DNA-binding domain to recognize any desired 15 base pair sequence in the genome in rapid fashion. The...All you do to create a protein that binds those 15 base pairs is take four plasmids out of the library in...genome-editing system for generating human stem cell-based disease models." Cell Stem Cell. 12, 238-251 (7 ...

...and what work he's planning on doing with CRISPR-based gene drives focusing on safety and the ability to...Board with a particular interest in generating Cas9 based tools to enable facile control of DNA and RNA.   ...

... knowledge base is going to necessarily need a diverse group of people to provide that base. Sometimes... Mission accomplished! One example of a mission-based team in academia would be your school’s post-doc...Let’s think about some of the advantages of team-based work: Nobody knows everything anymore John F. Kennedy... post, so here are just a few critical features based on my own experience: Leadership Few things are ...

...are functionally differentiated from one another. Based on their structural elements, oris interact with...proteins within different host organisms. pBBR1-based plasmids and plasmids in incompatibility groups ...campestris pv. malvacearum, Zymomonas mobilis pBBR1 based pBBR1  Gram-negative  Alcaligenes eutrophus...constitutive and IPTG-inducible integron promoter-based expression systems for Escherichia coli and Agrobacterium...

...infectious titer of a viral vector stock can vary based on transduction target and can also be altered by...titer is typically calculated by two popular PCR-based methods: Quantitative PCR (qPCR) Digital droplet...not widely used today (Fagone et al., 2012). PCR-based methods are robust, easy, fast, and convenient. ...quantified with a different primer pair. Ct values vary based on the amount of starting material in the PCR because...

... et al) we were very excited to set up a CRISPR-based editing platform in our lab. Our goal was to use...binding sequence, we recommend changing several bases as close to the PAM as possible (see below to find...Ludwig-Maximilians-University Munich. His lab develops iPSC-based models of neurodegenerative and neurovascular diseases...of Alzheimer’s disease by developing novel iPSC-based model systems.     References 1. Cong, Le, et al...

...of the desired mutation (usually 1-3 mismatched bases) is sufficient for your primers to successfully ...a minor extension can usually ensure that the 3’-base(s) do not form secondary structures. The introduction...feature used in TA-cloning). Such non-complementary bases at the 3’-end interfere with plasmid reconstitution...et al. "Predicting DNA duplex stability from the base sequence." Proceedings of the National Academy of...

...the best possible results for any tissue culture-based experiment, labs should routinely test for mycoplasma...detector cell lines, ELISA, fluorometry, or PCR-based approaches to detect even low levels of contamination...detection, Uphoff et. al. outline a do it yourself PCR-based approach complete with freely available positive...concentration and titration of pseudotyped HIV-1-based lentiviral vectors."Nature protocols 4.4 (2009):...

... decisions based on experimental timelines and specific aims. 2. Then make decisions based on financial...instructions on creating a budget, making decisions based on that budget, and negotiating in regards to the...will need, and how long it will take to publish based on these findings. Most often we suggest estimating...

...of imaging data (ILASTIK), or the popular plugin-based image process framework FIJI are just three of the...to the biological protocol, and assembly designs based around the use of commercially available tools and...provides a potential conflict between traditional IP-based models and the use of taxpayer money to support ...

...allows classification of individual target molecules based on their mutation status. The drop-off probe is ...References 1. Miyaoka, Y. et al. Isolation of single-base genome-edited human iPS cells without antibiotic..., Berman, J. R. & Postovit, L.-M. A Digital PCR-Based Method for Efficient and Highly Specific Screening...

...contributed by guest blogger, Rachel Rubinstein, a field-based software product manager at Oxford Nanopore Technologies... messy if you’re not on top of it. In a science-based software PM role, having a good understanding of...Rubinstein! Rachel Rubinstein is currently a Field-based Product Manager at Oxford Nanopore Technologies ...

... efficiencies ranking as low, medium or high is based on the reprogramming efficiencies reported in González...Non-replicating Plasmids: Generating iPSCs with plasmid-based expression requires serial transfection of 1 or ...Replicating oriP/Epstein-Barr nuclear antigen-1 (EBNA1) based episomes: These plasmids carry the origin of replication...DNA methylation. Another downside of using EBNA1 based vectors is that the plasmid can hang around for ...behave. iPSCs are typically tested with molecular-based assays first. These methods less rigorously assesses...Transgene Expression Mediated by a Sendai Virus-Based Vector Using a Nonstructural C Protein and Endogenous...

... of DNA you need, and at which level of purity. Based on these premises we can classify a plasmid preparation...will adopt for the extraction. Restriction enzyme based approaches can require several hundreds of nanograms...