Dr Nick Oswald's Profile

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Antibiotics Used in Molecular Biology

Antibiotics are used in a wide range of techniques in molecular biology. My aim with this post is to provide an easy reference to some of the main ones used in molecular biology, their mechanisms, range and working concentrations. I hope you will find it useful. Your Personal Antibiotics Reference Guide *Abbreviations: Gm(+/-)=Gram positive/negative; My=mycoplasma; […]

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The Essential PCR Troubleshooting Checklist

Routine PCR? Let’s be honest, there’s no such thing. Even with the simplest PCR reaction things can go wrong, so you need to have a good checklist of ideas for PCR troubleshooting and rectifying the problem. Today I have brainstormed all of the ways I can think of to approach problems with standard PCR reactions. […]

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In PCR, qPCR and qRT-PCR 27th of July, 2016
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E.coli Electroporation vs Chemical Transformation

This is the first in a three part series on the transformation of E.coli. By the end of this you should be an expert on E.coli transformation and on which strains to choose for different applications. If you’re already an expert, I hope it’ll be an enjoyable refresher for you. In either case, please comment […]

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How SDS-PAGE works

What does a stacking gel do? This and other questions about SDS-PAGE answered!

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The Basics: How Phenol Extraction of DNA Works

Phenol extraction is a commonly used method for removing proteins from a DNA sample, e.g. to remove proteins from cell lysate during genomic DNA preparation. It’s commonly used, but not commonly understood. If you want to know how it works so you can show off to all of your friends… read on. The basic protocol […]

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How to Shut Off Background Lac Expression in LB

Here’s a tip that you may find useful if you are expressing proteins in E.coli using a lac promoter-based expression system, e.g. pET, in LB medium (L-broth). Lac expression systems are typically induced in the lab using IPTG (isopropyl-beta-D-thiogalacto- pyranoside), which is a non- hydrolyzable analog of lactose, the natural inducer of the lac operon. […]

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PCR problems read on

PCR Problems? Try an Additive

You’ve tried all the usual stuff, and checked the primer sequences twice, but still can’t get that PCR fragment amplified. It’s time to enter the strange world of PCR additives. Over the years a variety of additives have been shown to enhance PCR reactions in certain situations. Here is a summary of some of the […]

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In PCR, qPCR and qRT-PCR 9th of July, 2016
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Perfectionism: Are you on the downward spiral?

Do you fear failure every time you do an experiment? Do you feel constantly stressed about obtaining poor results? Do you feel personally culpable when an experiment goes wrong? If you answered “yes” to any or all of these questions, you may be suffering from perfectionism. For a scientist, this is a particularly damaging trait […]

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In Personal Development 9th of July, 2016
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optimal temperatures read on

Why Do Enzymes Have Optimal Temperatures?

Every biologist is familiar with the profile of the rate of an enzymatic reaction versus temperature as shown in the figure. We know that enzymes from E. coli or warm-blooded animals tend to have an optimum around 37°C, while those from thermal vent bacteria have much higher optimal temperatures. Surprisingly, I find that many biologists […]

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Get Your Clone 90% Of The Time with Ligation Independent Cloning

Are you stuck in cloning hell?, Tired of doing ligations that don’t work? Want a faster, more efficient cloning procedure? You should try ligation independent cloning. A growing number of researchers swear by ligation independent cloning methods because they are simpler and more efficient than conventional cloning and as a recent convert to their ranks, […]

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