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The Fundamentals of Sample Preparation for Next Generation Sequencing

What you will learn in this webinar: Methods of extraction. Details on sample quality and quantity required for NGS and how to achieve this. Library preparation. Handy tips and tricks for library preparation. Summary: This webinar will detail sample preparation for Next Generation Sequencing (including methods of extraction, sample quality and quantity, and library preparation).…

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Best Practices for DNA Shearing for NGS

Construction of high-quality sequencing libraries is pivotal to successful NGS, and DNA quality is one of the most critical aspects of library preparation. As this Nature Methods paper illustrates, DNA shearing involves appropriate and consistent fragment sizes for sensitive and accurate sequencing, and the fragments must be accurately analyzed prior to sequencing to measure molarity…

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Probability Theory and Molecular Barcodes

In biology, a molecular barcode is a characteristic DNA sequence used to distinguish and gather together similar items. Such a simple but powerful concept is useful in various applications. As an example, the Barcoding of Life project aims to identify specimens through the sequencing of standard gene regions, and use these as barcodes. On the other…

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The 31 Flavors of Drosophila Electrophysiology Recording Solutions

If you want to know what is going on in the brain of drosophila you can use neurobiology imaging techniques to get a global whole-brain perspective. However, such techniques are slow compared to the rapid nature of the neuronal electrical activity, which may be better studied using Drosophila electrophysiology. Lab techniques are often shrouded in…

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If You Use Plasmid Midi and Maxi preps, Your Life is About to Get Much Easier

The plasmid midi and maxi preps are essential for generating high purity plasmid stocks for your precious experiments. Unfortunately, they are also tedious, time consuming, temperamental, and sometimes, utterly soul-destroying. Well, until now that is, because you are about to witness a massive change in the world of large-scale plasmid preps. And by the end…

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Troubleshooting RNA Isolation

As widely used as it is, isolating RNA remains one of the more finicky protocols. Just about anyone who has performed the technique has their own personal tips and tricks to successfully isolate intact RNA from their samples with consistency. Although RNA can be somewhat unpredictable since it is so labile, there are a few…

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How to Culture Primary Human Bronchial Epithelial Cells

Human bronchial epithelial cells (HBECs) are a challenge to culture. As highly specialised cells that exist in carefully ordered multi-layered structures, they are especially fickle and finding optimum conditions to keep them happy is tricky. The cultures are also extremely sensitive to tiny changes in routine or environment. However, there are certain basic principles that…

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Where did my DNA go? Tips on improving DNA Clean-Up

Genomic DNA clean-up is a technique that is very common but still causes many people to suffer from separation anxiety.  Here’s a look at some tips and tricks to improve your DNA clean-up and avoid the loss of precious samples. Why is my DNA “dirty”? Here’s the scene: You’ve collected a set of samples, they could…

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Some Sanger Sequencing Tips and Tricks

Sanger sequencing is still a workhorse of most molecular biology labs. Even with the advent of next-generation sequencing we still need to sequence our clones and PCR products. In this article I have listed some of the tips and tricks we used in our Sanger services. (1)Dilution of BigDye: I’d expect this to be a…

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Three Insights for Choosing the Best DNA Size Selection Method for Your Project

With so many methods for DNA size selection, it can be confusing to determine which is best for your project. Fortunately, academic researchers have evaluated many DNA size selection methods for a range of applications, and their conclusions provide some useful insight for the community. Double-Digest RAD-Seq Based on the original protocol for restriction site…

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How to Set up Your Elution Experiment

What do DNA mini preps and protein immunoprecipitation experiments have in common? They start differently, but they end with the same, critical stage – elution. But what exactly is elution, and what is the point? The Terminology First, let’s start with some basic terminology: Elution – extracting one material from another by washing with a…

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RNA-seq: The Challenges to Diving Right In

It’s the hot new technique. With a single procedure, you can get information about all RNA transcripts at once! It sounds like a dream. While RNA sequencing (RNA-seq) has opened the door to exciting new questions, scientists interested in pursuing this technique should be aware of the roadblocks ahead of them. While RNA-seq can be…

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Sequencing-by-Synthesis: Explaining the Illumina Sequencing Technology

The “sequencing-by-synthesis” technology now used by Illumina was originally developed by Shankar Balasubramanian and David Klenerman at the University of Cambridge. They founded the company Solexa in 1998 to commercialize their sequencing method. Illumina went on to purchase Solexa in 2007 and has built upon, and rapidly improved the original technology. Millions of reactions and…

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Why Is It Important To Run Your NGS Gels Consistently?

This article discusses some of the important things to consider if you are using agarose gel electrophoresis for size-selection of your NGS libraries. Gel electrophoresis is a simple and very commonly used technique in most labs. Careful! It’s a critical step However this simplicity means people can often overlook the fact that there are applications where…

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Using PubCrawler: How to Speed Up Your Literature Searches by Crawling

It often seems like a full-time job searching PubMed for the latest papers on adult neurogenesis, mutations in kinase genes that lead to cancer, or the newest papers published from a major figure in your field. However, falling behind on the latest published research can also be a significant source of stress. In search of…

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