title: PCR analysis of chloroplast double-strand break (DSB) repair products induced by I-CreII in Chlamydomonas and Arabidopsis.
abstract: Homing endonuclease I-CreII has been used to study the consequences and repair of a double-strand break (DSB) in the chloroplast genome of Chlamydomonas and Arabidopsis. Since I-CreII is from a mobile psbA intron of Chlamydomonas, it cleaves the psbA gene of an intronless-psbA strain of Chlamydomonas. And it cleaves specifically in the psbA gene of Arabidopsis, which is naturally intronless. We have shown further that most of the repair products of an I-CreII-induced break in chloroplast DNA can be defined by PCR analysis with total nucleic acids and the appropriate primers. Here, we provide protocols for small-scale preparation of nucleic acids from Chlamydomonas and Arabidopsis, as well as guidelines for the subsequent PCR analysis.
authors: Section of Molecular Cell and Developmental Biology, Institute for Cellular and Molecular Biology, School of Biological Sciences, University of Texas at Austin, Austin, TX, USA.
authors: Taegun
authors: T
authors: Kwon
authors: Obed W
authors: OW
authors: Odom
authors: Weihua
authors: W
authors: Qiu
authors: David L
authors: DL
authors: Herrin
mesh_terms: D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D002696:Chlamydomonas / Q000235:genetics* / Q000378:metabolism; D002736:Chloroplasts / Q000235:genetics* / Q000378:metabolism; D053903:DNA Breaks, Double-Stranded; D004260:DNA Repair; D018742:DNA, Chloroplast / Q000235:genetics* / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods
chemical_list: D018742:DNA, Chloroplast; D015252:Deoxyribonucleases, Type II Site-Specific