Finding your knockout: reverse genetics techniques for plants.
The process of finding a mutant plant for your gene of interest has recently become far more straightforward and painless than has ever been possible before. This has come about through the production of large-scale insertional mutagenesis populations. These can now be readily screened in bulk for insertional mutants through a mixture of molecular (PCR/hybridization) and bioinformation techniques. Here we describe a step-by-step guide to the molecular protocols and a description of the bioinformatics approach.
May
Sean T
ST
Nottingham Arabidopsis Stock Centre, School of Biosciences, Plant Science Division, University of Nottingham, UK. Sean_May@nasc.nott.ac.uk
Clements
Deborah
D
Bennett
Malcolm J
MJ
D000465:Algorithms; D017360:Arabidopsis / Q000235:genetics; D004251:DNA Transposable Elements / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D017343:Genes, Plant / Q000502:physiology*; D005821:Genetic Techniques; D008958:Models, Molecular; D016254:Mutagenesis, Insertional / Q000235:genetics / Q000379:methods*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; C036483:T-DNA