Nick Oswald

I started Bitesize Bio on a Macbook on my kitchen table in 2007 while in my 7th year of working as a molecular biologist in biotech. My aim was to share the know-how that I had acquired from the school of hard-knocks in the lab, so that others could learn from my mistakes and small victories. Nowadays my mission is to facilitate the gathering of hardcore know-how from whole spectrum of bioscientists and share it here on Bitesize Bio to create a super-mentor that any bioscientist can turn to for much-needed guidance.

Articles by Nick Oswald:

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…

09 Jul 2016 Protein Expression & Analysis

5 Tips on Vector Preparation for Gene Cloning

One of the most crucial steps in any cloning procedure is the preparation of the vector. Get it wrong and your chances of success will be drastically reduced. The overall aim for a good vector preparation is to obtain a fairly concentrated stock of undamaged, fully digested plasmid DNA that is free from contaminants. Missing…

09 Jul 2016 DNA / RNA Manipulation and Analysis

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,…

09 Jul 2016 DNA / RNA Manipulation and Analysis

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.…

09 Jul 2016 Protein Expression & Analysis

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…

09 Jul 2016 PCR, qPCR and qRT-PCR

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…

09 Jul 2016 Personal Development

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…

09 Jul 2016 DNA / RNA Manipulation and Analysis

Non-specific Binding? Tips to Sharpen up Your Western Blot

In the previous installment of this series on western blotting, we addressed potential sources of error when your final product is completely bare. But alternatively, what do you do when too much background is the problem? You may have beautiful bands of interest—but if there is a bunch of non-specific binding, your quantification and data…

15 Mar 2016 Protein Expression & Analysis

5 Ways to Clean Up A DNA Sample

One of the most common tasks in molecular biology is cleaning up DNA from aqueous solutions to remove buffer salts, enzymes or other substances that could affect downstream applications. Examples include cleaning up PCR reactions, digests or other enzymatic treatments and cleaning up genomic or plasmid DNA contaminated with cellular proteins/debris. There are several ways…

09 Jun 2015 DNA / RNA Manipulation and Analysis

Fast-track your Ampicillin Plasmid Transformations

Most of us use pretty standard transformation protocols for E.coli. Yours probably goes something like this: – Thaw the competent cells on ice – Add DNA – Electroporate (or incubate then heat shock for chemically competent cells) – Add rich medium (LB or SOC) – Incubate at 37°C (or appropriate temperature) for 30-60 minutes –…

19 May 2015 DNA / RNA Manipulation and Analysis