Experimental Techniques Explained

Click on the headings to view the whole articles.

Isothermal Titration Calorimetry

Isothermal Titration Calorimetry, or ITC, measures a very basic interaction- how does one thing bind to another? Imagine you have a protein, and want to measure how well it binds to its substrate. ITC allows you in a single experiment to gain a huge amount of thermodynamic information.

Shedding Light on Neuronal Networks

Neuroscientists now have a new tool capable of studying the function of particular groups of neurons through genetic manipulation followed by exposing cells to bursts of a particular wavelength of light.  The technique, called “optogenetics” allows precise manipulation of specific neurons that associate to particular functions through either stimulation or inhibition of electrical signalling within those cells.

Polymerase Chain Reaction (PCR)

Since its invention in the mid-1980s, the polymerase chain reaction (PCR) has been the cornerstone of molecular biology and genetics – almost like a scientist’s bread and butter! PCR acts like a molecular photocopier, producing huge numbers of identical DNA fragments from the original sample. The PCR products, identical to the original target sequence, can then be used to analyse the sequence and identify, for example, mutations in the amplified gene/region.

MALBAC- Whole Genome Amplification

MALBAC stands for Multiple Annealing and Looping Based Amplification. It improves upon previous whole-genome amplification methods by looping the full amplicon (the replicated DNA) such that it itself cannot be amplified reducing amplification bias (where one or multiple stretches of the genome are amplified to a greater extent than others). This leads to the production of DNA fragments from across the whole genome (a higher percentage coverage) and leads to accurate sequencing of a very small amount of starting DNA.

SDS-PAGE

Sodium Dodecyl Sulfate-Poly Acrylamide Gel Electrophoresis, commonly known as SDS-PAGE, is a very common technique used in the lab. It is used for the rough analysis of dirty (raw, impure) samples, sample separation and as a rough guide to protein quantity and identification. When used more carefully the technique can also be used to highlight protein weights, and thus roughly the lengths of the amino acid sequences that make up all proteins. But how does this experimental method work? Let’s find out!

Thin Layer Chromatography (TLC)

Thin layer chromatography, or TLC, is a technique used for the separation and analysis of molecules in a sample (Note- NOT DNA!). It can be used on amino acids, although in my lab it has been used to analyze the degradation (or lack of!) of large polymeric sugars by whole metabolically inactivated cells or by simple enzymes. Because of this, this article will be focusing on the analysis of saccharides (sugars).

Fluorescence Microscopy

This technique is actually really simple, and is based upon the principle that when you shine high energy light onto certain substances they will absorb and emit this light at different wavelengths.  If you can attach one of these fluorescent substances to whatever you’re trying to visualise, then you can look at it via a microscope.

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