天天操夜夜拍丨国产精品超清白人精品av丨天天爽天天噜在线播放丨国产精品久久久久久久福利竹菊丨色综合天天天天做夜夜夜夜做丨国内自拍xxxx18丨白人と日本人の交わりビデオ丨午夜激情在线观看丨成人18视频丨久久66热人妻偷产精品丨视频一区二区三区在线观看丨操操操插插插丨久久的久久爽亚洲精品aⅴ丨91精品一区二区三区在线观看丨aaa女人18毛片水真多丨99re在线视频精品丨超碰免费在丨中文字幕人妻无码专区app丨午夜香蕉视频丨又大又粗欧美黑人aaaaa片丨久久夫妻视频丨乌克兰性欧美精品高清丨亚洲欧美福利视频丨少妇高潮久久久久久一代女皇丨97国产高清

Tester for Chromosome XV

Before using any of the strains for mapping an unknown gene, check the strain with a known marker located on the specific chromosome for which the strain is being used. There have been reports of two incorrectly marked strains obtained from YGSC and from the Sherman-Wakem lab.

Mapping strains: chromosomal assignment with 2-micron tester strains (AB2)

Each of the 2-micron strains contains derivatives of the integrated plasmid YEp24 and is URA3+. It has become apparent that several of the 2-micron strains tend to lose the plasmid spontaneously at a rather high rate. To prolong stability of the integrated plasmid, revive the strains from paper replicas on Ura- medium rather than on rich YEPD medium. A study by Y. Kaneko of the Institute for Fermentation in Osaka, Japan, has established approximate percentages of loss of the Ura+ phenotype in the 2-micron strains when replica-plated from YEPD medium to Ura-. The results indicate that strains B-7588, B-7589, B-7175, B-7595, B-7596, B-7599, B-7602, B-7604, B-7608, B-7610, B-7612, and B-7614 become auxotrophic for uracil at a rate between 10 and 39%; the percentage loss of the Ura+ phenotype for all other strains is between 0.22 and 0.49%. For more precise numbers, please contact YGSC.

Temperature: 25°C

Atmosphere: Typical aerobic

Protocol: Protocol for the use of 2-micron tester strains:The method relies on a set of 16 [cir0] tester strains, each containing plasmid DNA integrated at or near the centromere of a different chromosome. The plasmid DNA derived from YEp24 contains the URA3+ gene, the 2-micron inverted repeat sequence, pBR322 sequences, and different yeast segments corresponding to centromeric regions of each of the 16 chromosomes. The 2-micron plasmid DNA remains stably integrated in [cir0] strains since the plasmid DNA lacks the 2-micron FLP gene required for site-specific recombination and the [cir0] cells contain no resident 2-micron plasmid to provide FLP function. Specific loss of integrated 2-micron plasmid DNA and the chromosome into which integration occurred is induced in a [cir0]/[cir+] diploid since FLP function provided by the 2-micron circles of the [cir+] parent catalyzes a site-specific recombination event. This results in the loss of the integrant plus the entire chromosome containing the integrant if the integration occurred at the centromere. The recessive mutation, in a [cir+] strain, can be assigned to its chromosome by crossing to each of the [cir0] tester strains and isolating the [cir0]/[cir+] diploids on selective medium. A subclone of each diploid is resuspended and plated on nonselective medium. Several hundred colonies of each diploid are replica-plated onto a medium that selects for the expression of the unmapped mutation. Each of the [cir0]/[cir+] diploids will lose the 2-micron plasmid DNA as well as the chromosome which contains the integration at a high frequency. The recessive mutation will be manifested only in the diploid containing the [cir0] tester strain in which the integrant is on the same chromosome as the mutation.

Protocol: Protocol for the use of 2-micron tester strains:The method relies on a set of 16 [cir0] tester strains, each containing plasmid DNA integrated at or near the centromere of a different chromosome. The plasmid DNA derived from YEp24 contains the URA3+ gene, the 2-micron inverted repeat sequence, pBR322 sequences, and different yeast segments corresponding to centromeric regions of each of the 16 chromosomes. The 2-micron plasmid DNA remains stably integrated in [cir0] strains since the plasmid DNA lacks the 2-micron FLP gene required for site-specific recombination and the [cir0] cells contain no resident 2-micron plasmid to provide FLP function. Specific loss of integrated 2-micron plasmid DNA and the chromosome into which integration occurred is induced in a [cir0]/[cir+] diploid since FLP function provided by the 2-micron circles of the [cir+] parent catalyzes a site-specific recombination event. This results in the loss of the integrant plus the entire chromosome containing the integrant if the integration occurred at the centromere. The recessive mutation, in a [cir+] strain, can be assigned to its chromosome by crossing to each of the [cir0] tester strains and isolating the [cir0]/[cir+] diploids on selective medium. A subclone of each diploid is resuspended and plated on nonselective medium. Several hundred colonies of each diploid are replica-plated onto a medium that selects for the expression of the unmapped mutation. Each of the [cir0]/[cir+] diploids will lose the 2-micron plasmid DNA as well as the chromosome which contains the integration at a high frequency. The recessive mutation will be manifested only in the diploid containing the [cir0] tester strain in which the integrant is on the same chromosome as the mutation.

ATCC <<--YGSC<<--L.P. Wakem and F. Sherman

Before using any of the strains for mapping an unknown gene, check the strain with a known marker located on the specific chromosome for which the strain is being used. There have been reports of two incorrectly marked strains obtained from YGSC and from the Sherman-Wakem lab.

Each of the 2-micron strains contains derivatives of the integrated plasmid YEp24 and is URA3+. It has become apparent that several of the 2-micron strains tend to lose the plasmid spontaneously at a rather high rate. To prolong stability of the integrated plasmid, revive the strains from paper replicas on Ura- medium rather than on rich YEPD medium. A study by Y. Kaneko of the Institute for Fermentation in Osaka, Japan, has established approximate percentages of loss of the Ura+ phenotype in the 2-micron strains when replica-plated from YEPD medium to Ura-. The results indicate that strains B-7588, B-7589, B-7175, B-7595, B-7596, B-7599, B-7602, B-7604, B-7608, B-7610, B-7612, and B-7614 become auxotrophic for uracil at a rate between 10 and 39%; the percentage loss of the Ura+ phenotype for all other strains is between 0.22 and 0.49%. For more precise numbers, please contact YGSC.