Roadside Assistance: Fixing Your Broken-Down ELISA

Roadside Assistance: Fixing Your Broken-Down ELISA

The ELISA (enzyme-linked immunosorbent assay) is arguably one of the most important and versatile tools in the toolbox of molecular biologists, biochemists and diagnosticians across the world. Defined by its simplicity and speed, the assay is easy to learn and perform in as few as five steps. But with so few variables to manipulate, an…

Express yourself:  Gene Co-Expression in Bicistronic Constructs

Express yourself: Gene Co-Expression in Bicistronic Constructs

Let’s say you work with a gene, and it has wonderful potential. Excitedly, you throw your gene into the cells and voila! It’s there. Great. Now what? Genes are powerful tools for directing cell activity, but thanks to that curiosity characteristic of scientists, we want to know more: So what do you do? You had…

The Art of Protein Expression – How Changes in the Universal Genetic Code Can Affect The Activity of Your Expressed Protein

The Art of Protein Expression – How Changes in the Universal Genetic Code Can Affect The Activity of Your Expressed Protein

Protein expression is an art. There are many routes to optimize a protein expression protocol, such as using different expression systems (e.g. E. coli, yeast cells, insect cells) or changing the expression vector or culture media for the expression host. Fortunately, optimizing the parameters mentioned above often leads to improvements in your protein expression results. However, there is one other…

drug discovery to development

Using ADMET to Move Forward from Drug Discovery to Development

Anyone who is remotely associated with any kind of drug development program knows how challenging the field is. Not only is the process complex and time consuming, but it also requires the concerted effort of experts from various disciplines such as chemistry, biology, microbiology, pharmacology, toxicology, etc. Whenever such collaborations are not possible, you are…

To boil? Or be boiled?  Saving Time With Colony PCR

To boil? Or be boiled? Saving Time With Colony PCR

Applying molecular techniques to unicellar organisms leads to many questions… All these questions can be answered by going through the slow, routine pipeline process in a molecular biology laboratory: Culture cells, harvest them, extract DNA and PCR. Or guess what?! You can directly perform PCR using the colonies on the petri dish that the post-doc…

Detecting Signal in qPCR: From DNA Binding Dyes to BHQ Probes
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Detecting Signal in qPCR: From DNA Binding Dyes to BHQ Probes

A Brief History of Detecting Amplicons The Old Days of Ethidium Bromide In the early 1990s, quantitative (q) PCR was in its infancy, and despite PCR itself already being around for 10 years, there were no easy ways of precisely quantifying the amount of DNA that was amplified in a PCR reaction. In those days PCR…

Fast-track your Ampicillin Plasmid Transformations

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– Spread onto antibiotic plates That…

Capillary Gel Electrophoresis: An Alternative to SDS-PAGE?

Capillary Gel Electrophoresis: An Alternative to SDS-PAGE?

When you think about separating proteins, do you think about separating them using a gel? Specifically using SDS-PAGE? If you answered “yes”, it is for good reason. SDS-PAGE is ubiquitous in molecular biology labs because it is good at separating proteins. However, SDS-PAGE takes a lot of time and is labor-intensive. So let’s expand your…

A Guide to Flow Cytometry Software: Becton Dickinson’s ‘Diva’

A Guide to Flow Cytometry Software: Becton Dickinson’s ‘Diva’

If you use a Becton Dickinson (BD) cytometer in your lab, the chances are you are acquiring your data using ‘Diva’ software. Diva software is used to acquire your cytometry data on LSRII, LSRFortessa, CantoII and Aria cell sorters. As well as acquiring your data using Diva software, you can also analyse your data after…

Seamless Ligation Cloning Extract (SLiCE) Explored and Explained

Seamless Ligation Cloning Extract (SLiCE) Explored and Explained

Traditionally, if you’re hoping to clone a DNA/RNA fragment (or insert) into a vector, such as a BAC you would need: This process looked something like this: cut you vector using exonucleases to expose ends that are complementary to the ends of your insert and then stick your insert into the vector using a ligase….

Heat Inactivation of Serum for Tissue Culture – Is it Necessary?

Heat Inactivation of Serum for Tissue Culture – Is it Necessary?

In the cell culture practice, heat inactivation of serum products has always been accepted and is one of the basic protocols passed on to new cell culturists. There is no strict standard protocol for heat inactivation, some say incubate at 56 °C for 30 minutes, while some say it can be efficiently performed using temperatures…

How to Troubleshoot Problems with Fluorescently Tagged Proteins
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How to Troubleshoot Problems with Fluorescently Tagged Proteins

Using fluorescent proteins as imaging probes is a widespread and versatile technique in microscopy. You can use them in a wide range of living systems, from single cultured cells to complete organisms and animals. Fluorescently tagged proteins can be used to track and examine real-time localization, interactions, and translocation of your protein of interest, as…

GeneDig: An Easy-to-use Genome Browser for Bioscientists

GeneDig: An Easy-to-use Genome Browser for Bioscientists

The recent advancement of next generation sequencing technology and the development of novel gene editing tools, such as CRISPR-Cas9, have revolutionized research in genetics. In this golden era of molecular biology, knowing how to dig and navigate through all the enormous sequence information is an essential skill for most molecular biologists. However, to obtain facile…

Top Tips on How to Prevent Cell Line Cross-Contamination

Top Tips on How to Prevent Cell Line Cross-Contamination

Recently we wrote an article about widespread cell culture contamination and how to detect it. This follow-up article will provide practical tips on avoiding cross-contamination in the first place. Be Cautious While Working The first way of cross-contaminating cultures is by accidentally mixing two cultures together, which may lead to an unintended co-culture or the displacement…

Live-Cell Imaging: Choosing the Right Technique
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Live-Cell Imaging: Choosing the Right Technique

If you want to see in real time what is going on inside your cell then you should be performing live-cell imaging. Live-cell imaging techniques allow real-time examination of almost every aspect of cellular function under normal and experimental conditions. With all live-cell imaging experiments, the main challenges are to keep your cells alive and healthy…

Genomic Analysis of Single Cells: The Benefits of Being Single

Genomic Analysis of Single Cells: The Benefits of Being Single

You don’t need to be told about how next generation sequencing technologies have revolutionized the way we study the genome and the epigenome. Whether you want to look at transcription (RNA-seq), translation (Ribo-seq) genomes (DNA-seq), interactions of proteins and DNA (ChIP-Seq) or to study epigenetic features such as methylation (whole genome bilsulfite sequencing) there are…

Catch of the Day: A Look into Different FISH Techniques

Catch of the Day: A Look into Different FISH Techniques

You are probably familiar with fluorescent in situ hybridization (FISH) to detect and localize the presence or absence of specific DNA sequences on chromosomes. But did you know there are numerous FISH experiment variations? Including high-resolution FISH and quantitative FISH? Read here about Fiber-FISH, Q-FISH, and Flow-FISH and decide if you would like to undertake one of…

From ChIP-seq to MeDIP: A Glossary of Different NGS techniques

From ChIP-seq to MeDIP: A Glossary of Different NGS techniques

It was not long since the commercialization of NGS (a little more than ten years ago) that scientists went beyond the basics and got creative with the new technology to study much more than just the sequence of DNA. In this article we highlight some of the different NGS technologies and methods available out there….

How to Be Greener – The Environmentally Friendly Guide to PCR

How to Be Greener – The Environmentally Friendly Guide to PCR

Science is an expensive business and those who use high energy-demanding techniques may not even realize just how expensive they are. The Cost of PCR Let’s looks at PCR. You need to pay for the machine, all the ingredients including expensive enzymes, a freezer and a fridge for your ingredients, tubes and caps, not to…

Am I Damaging My  E. coli by Spinning at High Speeds?

Am I Damaging My E. coli by Spinning at High Speeds?

Dear Aunt Yersinia, A very annoying postdoc in our group keeps telling me off for spinning E.coli at 13K in a tabletop centrifuge. The postdoc claims that high speed damages cytoskeleton and this will reduce my transformation frequency. But I don’t believe her as the cells are cushioned by water during centrifugation. Can you tell…