Phenol-chloroform extraction & EtOH precipitation protocol

Peccoud Lab Protocol: purifying and/or concentrating DNA/RNA from various sources by phenol-chloroform extraction and ethanol precipitation. Introduction Sometimes it becomes necessary to concentrate your DNA/RNA or obtain purer DNA/RNA samples. If your DNA is resuspended in nucease-free water, you can concentrate it just by drying it down in the  Vacuum Centrifuge . However, if the DNA [...]

Phenol-chloroform extraction & EtOH precipitation protocol2019-02-06T19:37:38+00:00

Primers for gene deletion in yeast (design protocol)

Peccoud Lab Protocol: how to design primers for generating a de novo gene deletion in yeast Introduction Yeast strains carrying single deletions of non-essential genes are commercially available from GE Life Sciences (http://dharmacon.gelifesciences.com/cdnas-and-orfs/non-mammalian-cdnas-and-orfs/yeast/yeast-knockout-collection/). However, these are available only as deletions in which the gene is replace with the kanMX selectable marker (G418 resistance). It is [...]

Primers for gene deletion in yeast (design protocol)2019-02-06T19:37:38+00:00

DNA extraction protocol for yeast genomes or plasmids

Peccoud Lab Protocol: Isolating genomic DNA or plasmids from yeast Introduction The following protocol will recover total DNA and, with the indicated modifications, is useful to recover plasmids or genomic DNA. For plasmid recovery, the total DNA is transformed into E. coli. For genomic DNA to be used for PCR amplification - such as testing for [...]

DNA extraction protocol for yeast genomes or plasmids2019-02-06T19:37:38+00:00

SDS-PAGE protocol: visualizing proteins on acrylamide gels

Peccoud Lab Protocol: visualizing proteins via sodium dodecyl sulfate-polyacrylamide electrophoresis (SDS-PAGE) Introduction Because cellular extracts contain thousands of different proteins at a wide range of concentrations, it is often difficult to detect and measure specific proteins in these mixes, even when proteins are expressed at high concentrations. The principles of protein gel electrophoresis are similar to [...]

SDS-PAGE protocol: visualizing proteins on acrylamide gels2019-02-06T19:37:40+00:00

Agarose Gel Electrophoresis

Introduction In agarose gel electrophoresis, a sample of a biological molecule, in this case, DNA, is loaded into a well formed in an agarose gel matrix. To prevent the DNA from diffusing out of the wells of the gel, a small amount of loading buffer containing glycerol is added to the DNA solution. The loading [...]

Agarose Gel Electrophoresis2019-04-15T16:56:54+00:00

FBI Synthetic Biology Training

FBI Training Objectives For one week in May, we hosted 11 agents and analysts from the U.S. Federal Bureau of Investigation for an intensive synthetic biology training program. The goal of this event was to give the law enforcement personnel foundational knowledge and insight into the rapidly evolving field of synthetic biology. At first glance, a [...]

FBI Synthetic Biology Training2019-02-06T19:37:41+00:00