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Plasmids 101: Blue-white Screening

Posted by Jessica Welch on Jun 4, 2015 9:03:00 AM

This post is part of our ongoing Plasmids 101 series. Plasmids 101 will provide you with an overview of general molecular biology knowledge and techniques. If you are interested in reading more, you can find the rest of the Plasmids 101 posts here.

Now that we have covered antibiotic selection here at Plasmids 101, we can talk about an even more specific method of screening your cloning reaction. Being able to select for colonies that contain your plasmid is a great start when cloning, but how about being able to choose those that contain plasmid with an insert? Blue-white selection is a widely used method to do just that!

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Topics: Plasmid Elements, Plasmids 101

Plasmids 101: Control Plasmids

Posted by Chari Cortez on Apr 9, 2015 11:29:00 AM

There are many, many different types of experiments carried out by scientists every day. Although the designs and outcomes may vary, one thing should be present in every experiment-based investigation of a hypothesis: proper controls!

For every experiment, an investigator needs a standard against which the results can be compared; results from an experiment lacking the proper controls are invariably inconclusive and unreliable. Proper controls provide the constant variables that enable the correct interpretation of the effect of the independent variable you are testing. Importantly, they demonstrate the functionality of your experimental system and help identify opportunities for troubleshooting or optimization within your experiment. Read on to learn more about the various controls that can be used for plasmid-based experiments.

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Topics: Lab Tips, Plasmids 101

CRISPR 101: A New Series on Genome Editing & CRISPR-Cas

Posted by Marcy Patrick on Mar 5, 2015 12:06:12 PM

I am sure by now you have heard of CRISPRs. (If not, you can get up to speed here and here and here.) With such a fast moving technology, it is sometimes hard to keep pace with the new advances let alone remember the (maybe) long forgotten details of the biological process required to effectively design and utilize these tools. We certainly understand and are here to help!

Starting next week, we'll release the first post in our newest blog series - CRISPR 101 - a companion series to our popular Plasmids 101 articles. These posts are created to educate all levels of scientists and provide a resource for some of the basic principles driving CRISPRs and genome editing technology.

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Topics: Genome Engineering, Plasmids 101, CRISPR, CRISPR 101

Plasmids 101: E. coli Strains for Protein Expression

Posted by Julian Taylor-Parker on Feb 10, 2015 10:06:00 AM

In a previous Plasmids 101 blog, we reviewed the salient features of several popular strains of E. coli for DNA propagation. While great for cloning purposes, these E. coli strains are not usually well suited for recombinant protein expression. Many challenges can arise when over-expressing a foreign protein in E. coli. We will review the potential pitfalls of recombinant protein expression and some of the most popular commercial strains designed to avoid them.

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Topics: Lab Tips, Plasmids 101

Plasmids 101: Cre-lox

Posted by A Max Juchheim on Jan 13, 2015 10:47:00 AM

In previous posts for our Plasmids 101 series, we examined a number of important plasmid elements – promoters, origins of replication, protein tags, and antibiotic resistance markers (just to name a few). In this edition, we’re going to take a look at a very interesting tool that can be used for creating (excuse the pun) specific, targeted DNA modifications in transgenic animals, embryonic stem cells, and/or tissue-specific cell types: Cre-lox recombination.

What is Cre-lox?

The Cre-lox system is a technology that can be used to induce site-specific recombination events. The system consists of two components derived from the P1 bacteriophage: the Cre recombinase and a loxP recognition site. The P1 bacteriophage uses these components as part of its natural viral lifecycle, and researchers have adapted the components for use in genome manipulation.

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Topics: Plasmid Technology, Plasmid Elements, Plasmids 101

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