Home > Protocols

We obviously haven't updated this page in a long time, but the following protocols may still be useful for those new to DNA/mRNA extraction and cloning or at least they show how the field has grown during the past decades:

DNA extraction protocol. We finally abandoned the traditional phenol-chloroform extraction in favor of a kit (yikes!) from Omega Biotek (E.Z.N.A. Mollusk DNA Isolation Kit). With this kit we get better extractions (based on the ability to amplify single copy nuclear genes) than with other kits or methods. The protocol listed here has a couple of slight modifications to the manufacturer's original protocol.

cDNA Prep protocol . This is an adaptation of various protocols for extracting mRNA and for producing cDNA that can be used as a template in PCR or for other procedures. It is quick (15 minutes of work and 90 minutes of incubations) and easy and relatively inexpensive (and so we like it very much!). While we typically use this protocol with Conus venom ducts, the approach can be used with other tissues and other organisms (it was initially developed for constructing cDNA from urchin gonads). This protocol was originally described by Duda & Palumbi (1999).

Cloning protocol. This is our modification of Invitrogen's TA cloning kit protocol. Our protocol utilizes lower volumes of reagents than recommended by Invitrogen which helps to substantially reduce cloning costs! The protocol is also written for new recruits to the clone wars.