Simplified primer design for PCR-based gene targeting and microarray primer database: two web tools for fission yeast.
PCR-based gene targeting is a popular method for manipulating yeast genes in their normal chromosomal locations. The manual design of primers, however, can be cumbersome and error-prone. We have developed a straightforward web-based tool that applies user-specified inputs to automate and simplify the task of primer selection for deletion, tagging and/or regulated expression of genes in Schizosaccharomyces pombe. This tool, named PPPP (for Pombe PCR Primer Programs), is available at http://www.sanger.ac.uk/PostGenomics/S_pombe/software/. We also present a searchable Microarray Primer Database to retrieve the sequences and accompanying information for primers and PCR products used to build our in-house Sz. pombe microarrays. This database contains information on both coding and intergenic regions to provide context for the microarray data, and it should be useful also for other applications, such as quantitative PCR. The database can be accessed at http://www.sanger.ac.uk/PostGenomics/S_pombe/microarray/.
Penkett
Christopher J
CJ
Cancer Research UK Fission Yeast Functional Genomics Group, Wellcome Trust Sanger Institute, Cambridge CB10 1HH, UK.
Birtle
Zoë E
ZE
Bähler
Jürg
J
D017931:DNA Primers / Q000737:chemistry / Q000235:genetics*; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics*; D020869:Gene Expression Profiling / Q000379:methods; D018390:Gene Targeting / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D012568:Schizosaccharomyces / Q000235:genetics*; D012984:Software
D017931:DNA Primers; D004271:DNA, Fungal