Techniques

Codon Optimization for Increased Protein Expression

articodon optimizations tools in synthetic biology

The Genetic Code: A Universal Template for Protein Translation All known organisms share the ‘central dogma’ of molecular biology. DNA is transcribed into mRNA that is translated into protein. During the discovery of the genetic code, Francis Crick hypothesized that translation required a mediator to aid mRNA-guided translation according to a number of specifics. Amongst…

Read More

Antibody Validation

antibody validation protocol and research

Not all antibodies are valid for every experiment and condition, and they must be validated for the specific application and species. Currently, there is no standard means of “antibody validation,” and this can greatly impact experimental reproducibility and reliability. Journals and granting agencies have taken steps to address this gap. Many now require you to…

Read More

How to Use CellProfiler for Cell Imaging

Are you trying to figure out how to calculate intensities of fluorescently-labeled single cells? Do you have cells at high densities or present in clusters? Are you worried that your current cell imaging analysis software is unable to mark clear boundaries around each cell in a cell cluster? Don’t fear, because CellProfiler 2.1 is here to…

Read More

Cre-loxP Recombination Essentials Part 2

The Cre-loxP recombination system is routinely used for the generation of mouse knockouts. In part 1 of this mini-series, I introduced the concept and applications of Cre-loxP. As with any other technology or research tool, it has limitations and pitfalls that need to be considered while planning experiments or interpreting results. This article will take…

Read More

A Crash Course in CRISPR-Cas9 Editing in Drosophila

CRISPR-Cas9 has become a magic tool for molecular biologists, transforming genetic engineering from a once unbelievable dream into tangible reality. Today, you can easily edit primary cells or cell lines within a few weeks with well-established protocols or others’ hands-on advice. However, no matter how many success stories you find online or hear at seminars…

Read More

Introduction to DREADDs – Control Over G Protein Coupled Receptor GPCR signaling

DREADDs

Gee, Protein, What Do You Do? Manipulation of a system under investigation is the backbone of experimentation. A new tool called Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) allows us to hijack cell signaling and study cell function within living organisms. Like its cousin technique, optogenetics, DREADD technology uses a viral vector to introduce…

Read More

Sheath Pressure: Nozzle Size Does Matter

sheath pressure

Hello again, fellow Flow Cytometry Fan! It looks like you have your experiment all planned out, including staining protocols and gating schemes, and are ready to get some paradigm-shifting data. But before we start “plugging-and-chugging” samples through your cytometer of choice, we need to make sure that the nozzle size and sheath pressure are set…

Read More

A Primer on Phage Display Libraries

phage display

Phage display – the process of genetically fusing antibody fragments with phage to identify binding partners to your protein of interest – was covered pretty thoroughly here over the past few months. The success of this assay predicates on creating a diverse library of up to 1012 genes coding for these antibody fragments. Despite being…

Read More