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Plasmids 101: Optimizing Plasmid Yields

Posted by Amanda Hazen on May 26, 2016 10:30:00 AM

This post was updated on March 21, 2018.

Most of the time, plasmid prepping is a breeze. You get your stab from Addgene, streak for single colonies, sub-culture, and prep with a DNA prep kit or your lab's favorite in-house protocol. DNA yields for this procedure are typically in excess of 100 ng/ul, more than enough DNA to verify your plasmid via sequencing or restriction digest.

Chances are, you’ll even have DNA left over for other applications, like PCR, cloning, transfection, or long-term storage. But what about those pesky situations where your plasmid yield is sub-optimal? If you are consistently getting sub-optimal plasmid yields from your prep, try our tips below!

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Topics: Lab Tips, Plasmids 101

The Challenges of Cell Culture

Posted by Guest Blogger on May 5, 2016 10:30:00 AM

This post was contributed by ATCC Outreach Scientist Nick Amiss.

Cell Culture in the Present Day

Cell culture is a widespread tool used in the fields of oncology, virology, immunology, microbiology, and pharmacology (many of which are represented in Addgene's Special Collections). Arguably, oncology has benefited most as cancer cells are amenable to culture in vitro. Virologists too have benefited from the ability to propagate viruses in cell culture; if there were no cells to infect, there would be no viable viruses to study. Interestingly, due to certain high profile problems surrounding cell culture over the last few decades some pharmacologists have tended to avoid cell culture in favour of biochemical assays followed by in vivo testing. The mantra being “you can’t trust those whacky cell lines”. This may certainly have been a valid concern for the discipline (or lack thereof) in times gone past but these days it’s never been easier to conduct high quality work in cell culture.

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Topics: Lab Tips

Getting the Most from Your Lentiviral Transduction

Posted by Meghan Rego on Apr 7, 2016 10:30:00 AM

Lentiviruses are a powerful laboratory tool often employed to establish cell lines that stably express a gene of interest. While the general approach for using lentivirus, infect and select, seems simple, in actuality, many find using lentivirus to be time consuming, difficult, and lacking in reproducibility. Read on for some tips for getting the most out of your lentiviral transduction experiments.

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Topics: Lab Tips, Viral Vectors

Addgene's Tips for Plasmid Quality Control

Posted by Julian Taylor-Parker on Jan 14, 2016 10:30:00 AM

One of the best things about sharing plasmids through Addgene is that we provide an added level of confidence in the plasmids we distribute through our quality control processes. Every plasmid we receive is rigorously verified before becoming available to the community.

This is no small task, however, at a repository that consistently receives around 200 new DNA samples every week. Here we will provide an inside look at the steps we take to verify the identity and quality of the plasmids we make available and provide some advice on the steps you can take to verify your own plasmids.

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Topics: Plasmid How To, Inside Addgene, Lab Tips

Tips from the Repository Trenches: Using Barcodes to Track Samples

Posted by Amanda Hazen on Oct 6, 2015 10:30:00 AM

Every plasmid sample enters Addgene the same way. A package is delivered by a mail courier and then the journey of transformation and storage begins. Some samples are submitted as bacterial colonies in petri dishes, but close to 80% of samples are received as DNA in a microcentrifuge tube. Once a sample enters Addgene’s lab, a series of events is triggered. Each step is tracked through barcoded tubes that are scanned into our Laboratory Information Management System (LIMS), a database we use to keep track of and manage all of the samples that come through our doors. It would be a nightmare if we didn’t have a simple way to track all of our 40,000 samples available for distribution!

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Topics: Inside Addgene, Lab Tips

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