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Plasmids 101: Multicistronic Vectors

Posted by Melina Fan on Sep 9, 2014 4:20:00 PM

Co-expression of multiple genes is valuable in many experimental settings. To achieve this, scientists use a multitude of techniques including co-transfection of two or more plasmids, the use of multiple or bidirectional promoters, or the creation of bicistronic or multicistronic vectors. Unlike promoters which will create unique mRNA transcripts for each gene that is expressed, multicistronic vectors simultaneously express two or more separate proteins from the same mRNA. We've discussed promoters before so in this blog post we’ll cover basics of multicistronic vectors: why they are useful, how they work, and how to get started with them.

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Topics: Plasmid Technology, Plasmids 101

A Conference By Postdocs For Postdocs: Future of Research

Posted by Joanne Kamens on Sep 2, 2014 11:05:00 AM

This post was originally published on LinkedIn. Follow Addgene on LinkedIn for repository news and updates.

Scientists must do science to be happy. What do we have to change to ensure that all scientists can have successful careers doing science in some form? There is a lot of talk about the state of scientist training in the US and around the world. There are rumors that we are training too many scientists and some propose radical changes to the way we view the graduate school and postdoc training years.

There is no doubt that there are too many scientists in the pipeline were they all to pursue jobs in academia. Certainly funding for academic research and training is getting tighter and competition is fierce. However, I believe there are plenty of great jobs out there for science PhDs. The problem is that too few of these trainees are sufficiently prepared during their 6-12+ (!) years of training to get jobs. They are not exposed to the vast non-academia career landscape and there is insufficient (or no) emphasis on developing transferable skills to enable pursuit of these diverse opportunities after training.

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Topics: Career

Plasmids 101: How to Verify Your Plasmid

Posted by Lianna Swanson on Aug 28, 2014 11:34:00 AM

Congratulations, you have a plasmid expressing your gene of interest (YGOI) and are ready to dive into your functional experiments! Whether you’ve cloned the plasmid yourself or obtained it from a colleague down the hall, it is always a good idea to take some time to confirm that you are working with the correct construct, and verify that the plasmid you received matches the expected sequence. Here at Addgene, we use NGS-based quality control to confirm the sequence of all the plasmids we distribute. This method is time-intensive, so we recommend two other methods for quick plasmid verification: Sanger sequencing and diagnostic restriction digest.

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Topics: Plasmid How To, Lab Tips, Plasmids 101

Advice for Moving Into Sales After Your Science Postdoc

Posted by Guest Blogger on Aug 21, 2014 11:15:00 AM

This guest blog post was contributed by Seán Mac Fhearraigh, PhD, of www.phdadvice.com.

I sold out, well in part that’s what leaving academia feels like for a lot of researchers. For many years I struggled with the questions of “Will I leave academia?” and "What type of science careers would would make a good fit for me?" Whatever would I do next and what options lie ahead of me? Like a considerable number of researchers, I had high hopes of securing tenure in a British or Irish University where I would continue on with my academic dream of studying cell division. However, after 6 years in the business of western blots and cloning, I decided to pack it in. I had just started month 8 of my 3 year Post-Doctoral contract at the University of Cambridge and knew it was time to leave. I really enjoyed my time in the lab, my colleagues and what I was researching, but I decided I needed a new challenge.

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Topics: Career, Career Readiness

Choosing Your Perfect Empty Backbone

Posted by Lianna Swanson on Aug 19, 2014 11:39:33 AM

Vectors (or empty backbones) are frequently used in molecular biology to isolate, multiply, or express the insert they carry in the target cell. These vectors allow you to test the function of Your Gene Of Interest (YGOI) in a controlled environment under various conditions. The first thing you'll need to decide when running your experiment, is which vector will best suit your needs?

At Addgene, we have a vast collection of empty backbones that have been designed, tested, and published by academic scientists. To help you find the vector that fits your experiments, I've described below some of the most frequently requested vectors in our repository and will discuss some of the features you may want to consider as you make your choice.

The first and most important thing you need to know is your expression system or environment. The host organism will determine the type of vector that you will need. You will also have to make sure that your plasmid has been incorporated into the host organism, usually achieved with the proper selection marker or antibiotic resistance.

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Topics: Plasmid How To, Plasmid Technology, Plasmid Elements

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