...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...

...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 ...

...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...

...Engineering Using CRISPR/Cas Systems has expanded out of control. To further consolidate these FAQs, Le Cong worked...gRNAs with the Cas9 nickase which will together introduce cleavage of the target site with efficiency similar...using CRISPR-Cas9 at the lab bench Q4: After the introduction of a mutation into the genome, how can cells...

... 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...

...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 ...

...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...

...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...

...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!...

...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 ...

...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...

...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 ...

...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...

...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...

...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...

...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...

...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...

...Promoters control the binding of RNA polymerase and transcription factors. Since the promoter region ...promoters can be regulated by positive or negative control. Positive inducible In the OFF state, the promoter...system, the activator rtTA (reverse tetracycline-controlled transactivator) is normally inactive and cannot...

...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 ...

...” and making sure we have adequate process and controls in place to manage those scenarios. What is the...the effectiveness of the Plan's risk management, control, and governance processes. Prior to Tufts Health...Auditors and the Information Systems Audit and Control Association. Lastly, Carl also serves on the Board...