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TypeBlog Post...and/or the use of chelating agents that can be introduced in the purification protocol to remove endonucleases... your preparation for a long time. Tris buffer controls the pH, while the EDTA chelates any divalent cations...of using a Tris buffer is that it allows better control of the pH preparation stabilizing the plasmid, ...
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Tips for Improving Your Next Manuscript
TypeBlog Post... sections of a paper, the abstract, figures, introduction, etc. so as to thoroughly explain the do’s and...reader is not in their head. The scientist has to introduce the reader to the manuscript and engage them in...to make the final product with an interesting introduction and concise discussion. Many thanks to our ... -
FastCloning: A Newer, Simpler Cloning Technique
TypeBlog Post...sequence the plasmid to confirm no mutations were introduced during PCR or any subsequent steps. Using high...DNA polymerases, I have never had any mutations introduced into my inserts. However, with any PCR-based ...PCR-based cloning technique there is a chance of introducing an unwanted mutation. Tips for high efficiency cloning... -
Management for Scientists: What Makes a Good Manager Anyway?
TypeBlog Post...recognition come from the successes of the team. A strong sense of engagement in the work Behind-the-scenes... This is the first in a 5 part series on Introduction to Managing People for Scientists. Subscribe...follow the entire series as it is published. Good introductory reference books on managing people: Skills ... -
Choosing Your Perfect Empty Backbone
TypeBlog Post...function of Your Gene Of Interest (YGOI) in a controlled environment under various conditions. The first...expression system and go for a lentiviral or a retroviral backbone. This is considered to be a more robust...great for overexpressing YGOI in an isolated, controlled environment. In most cases bacterial expression... -
History of CRISPR Cas - A tale of survival and evolution
TypeBlog Post... store a record of invading phages but to also destroy the phage upon re-exposure (2, 3). More specifically...complex binds to the foreign DNA, a cut is made to destroy the invader. Wikipedia, accessed 25 November...ultimate victor will be the species who has the strongest will to survive. Genome engineering and future... -
Arabidopsis in Education: How the Arabidopsis Biological Resource Center Brings Plant Science to Life
TypeBlog Post...researchers and educators in 60 countries each year. Introducing Arabidopsis Arabidopsis is an important model...develop a new teaching resource or you want to introduce Arabidopsis to your class for the first time –...Vectors at Addgene Find Simple Protocols for Introductory Lab Work MoClo Plants Parts Kit ... -
Antibodies 101: Epitope Availability
TypeBlog Post...cases, pay special attention to designing your controls in validation, as you may need to test different...great deal of frustration when designing and troubleshooting immunoimaging assays. It can also elucidate...Antibodies 101: Polyclonal Antibodies Antibodies 101: Introduction to Antibodies Resources on Addgene.org ... -
Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation
TypeBlog Post...One of the main tenets of our quality control (QC) mantra is to do our utmost to ensure that scientists...Gene Therapy. Existing methods for AAV quality control Historically, AAV vector QC has been sparse including...number of AAV preparations this level of quality control is sufficient but for viral vector core facilities... -
Plasmids 101: Origin of Replication
TypeBlog Post...the origin of replication (ori) and all of its control elements. The ori is the place where DNA replication...depending on how they are regulated. Generally, control of replication is referred to as "relaxed" or "...Vectors Copy Number+ ori Incompatibility Group Control pUC ~500-700 pMB1 (derivative) A Relaxed pBR322... -
Summer SciComm Series: Master of SciComm
TypeBlog Post...interests and areas of expertise. Perhaps the strongest aspect of science communication, then, is its ...address this obstacle in its many forms. A short stroll from my office to the tearoom would often illustrate...Cognitive LoadSummer SciComm Series: Modes of CommunicationIntro to Our Summer SciComm Series!... -
Another Pathway into Cells: iTOP
TypeBlog Post...problems. For instance, it’s much more difficult to introduce a gene of interest into primary cells, so most...both purified CPP-tagged protein and an untagged control could enter cultured cells and activate expression...reporters Previous Protein Delivery Techniques: Introduction of macromolecules into cultured mammalian cells... -
What Good Citizenship Can Do for Reproducibility in Science
TypeBlog Post...reproduce someone else’s result. Then, they hit trouble. The experiment won’t work. Even if it does, they...to share with other researchers to help them troubleshoot and understand. All of this communication and... of incentives. Right now, researchers are not strongly rewarded for being transparent in their communication... -
The Importance of High Titer for AAV Transductions
TypeBlog Post...therefore higher titer will be required. For example, retrograde AAV gets transported up the cell’s axon. While...even larger area, thus necessitating even higher retrograde AAV titers for successful transduction. In another...from and what do they mean? AAV Vector Quality Control: Going the Extra Mile with NGS AAV: Versatile ... -
Improving Transduction Efficiency with Exosome AAVs
TypeBlog Post...transduction profiles in different AAV serotypes both in vitro and in vivo when compared to the conventional AAVs...target less accessible cells, such as brain tumor stromal cells in a mouse model of glioblastoma (Volak et...vector-mediated genetic modification of brain tumor stromal cells after intravenous delivery." Journal of neuro-oncology... -
The Crispr Babies Saga Shows the Need for Action, Not More Delays
TypeBlog Post...independently verified, the watching world erupted in controversy. Briefly, there are two overlapping, valid critiques...within their respective mandates, to tackle a controversial and ethically fraught rapidly emerging technology...good research practice: computer simulations, in vitro research, animal models, and several stages of human... -
Plasmids 101: Mammalian Vectors
TypeBlog Post...components, it may be useful to describe the means of introducing genetic material (such as plasmids) into mammalian... comparable to bacterial transformation (the introduction of DNA into bacterial cells); however, the techniques...plasmid to be expressed transiently. Since the DNA introduced in the transfection process is not integrated... -
OCaMP: A New Calcium Indicator for Neural Imaging
TypeBlog Post...which is more than just the stuff that makes bones strong; it plays critical roles in nerve and muscle function...fluorescent tools are used simultaneously to monitor or control different cell types or signals in the brain. For...Simultaneous fluorescence (upper trace) and electrophysiological recordings (lower trace) using OCaMP expressed... -
GCE4All: Making Genetic Code Expansion Accessible
TypeBlog Post...acid in the media and the tRNAs and synthetase introduced to the bacteria using plasmids. Rapid expansion...plasmid. Third, a second plasmid will be needed to introduce the gene of interest containing the chosen codon...researchers, from selective protein ligation through introducing ncAAs with protein modifications of interest ... -
Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies
TypeBlog Post...this final installment, I’ll provide a brief introduction as to how you can engineer new transgenic flies...use attP sequence docking sites so that you can control exactly in which locus your transgene ends up (...selection markers include vermillion (another gene controlling eye color) and yellow (a gene affecting the body...