Chlamydomonas Center - Mapping Kit

An Online Informatics Resource for Chlamydomonas

	Genome Browser	Databases	Blast	Tools	Maps

HOME

Strains

Resources

Jobs

Educational

Links

Meetings

Methods

Newsgroup

KIT FOR MOLECULAR MAPPING OF CHLAMYDOMONAS GENES

These web pages are a resource for anyone interested in map-based cloning in Chlamydomonas reinhardtii. They are part of a mapping kit, which was designed to act as a starting point in the process of mapping a Chlamydomonas gene or mutation. The kit is intended to be a representative group of markers distributed as evenly as possible across the 17 linkage groups. This distribution of usable markers should allow the location of your gene of interest to be narrowed down quickly by screening each of the chromosomes at more than one position. The screening is accomplished with PCR and gel electrophoresis of a given mapping population's DNA, generated by crossing the mutant of interest (in mating type plus) with the polymorphic strain S1 D2 mt- (CC-2290, available from the Chlamydomonas Center)

The markers included in this kit were selected because an obvious size difference is seen for C. reinhardtii 137c (the usual lab strain) versus S1 D2 (a field isolate from Minnesota), making gel analysis simple. Each set of 2-3 primers was tested to determine whether they would be suitable for bulked segregant analysis, which reduces the number of PCR reactions required to map a gene. In cases where a marker is not suitable for BSA, individual DNA samples can still be readily screened. The web pages show how many DNA samples were mixed in a single tube, and how well the bulked DNAs performed. The availability of these gels on the web will permit anyone to see what was done for each marker along with the resulting gel, which allows each researcher to make their own decisions about which markers to use, and whether or not to bulk their samples.

Included in this kit are all of the primers needed for each marker for PCR. The primers are pre-mixed and diluted, and are distributed in a single 96-well plate. Each well contains enough primer for approximately 75 reactions. The kit also includes an instruction packet, comprising a key to the 96-well plate, a highlighted map of the linkage groups to show the overall distribution of markers in the kit, and a protocol for PCR and gel analysis.

Some examples of BSA, and a consolidated map of Chlamydomonas molecular markers, can be found in the Plant Physiology focus issue [February 2005, paper by Rymarquis et al.]. Those interested in ordering this kit or desiring more information should contact chlamy@duke.edu. There will be a nominal charge of $50 for the kit.