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Automated DNA sequencing is an extremely valuable technique which requires very high quality DNA templates to be carried out successfully. While it has been possible to readily produce large numbers of such templates from M13 or other single-stranded vectors for several years, the sequencing of double-stranded DNA templates using the ABI 373 DNA Sequencer has had a considerably lower success rate. We describe how the combination of a new fluorescent, dideoxy sequencing method, called cycle-sequencing, coupled with modifications to template isolation procedures based on Qiagen columns, makes fluorescent sequencing of double-stranded templates a reliable procedure. From a single five milliliter culture enough DNA can be isolated (up to 20 micrograms) to do 4-8 sequencing reactions, each of which yields 400-500 bases of high quality sequence data. These procedures make the routine use of double-stranded DNA templates a viable strategy in automated DNA sequencing projects. 相似文献
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Darryl L. York John L. Cummings Richard M. Engeman Kate L. Wedemeyer 《International biodeterioration & biodegradation》2000,45(3-4)
Bird strikes to aircraft are a serious economic and safety problem in the United States, annually causing millions of dollars in damage to civilian and military aircraft and the occasional loss of human life. We observed movements of 1236 neckbanded lesser Canada geese (Branta canadensis parvipes) to determine efficacy of hazing as a means to reduce goose presence at Elmendorf Air Force Base (EAFB), Anchorage, Alaska from August to October 1997. Emphasis was on movements of geese onto EAFB with additional data collected at the other two major airports in the area, Anchorage International Airport (AIA) and Merrill Field Airport (MFA). Daily observations indicated the presence of 208 individual neckbanded geese on EAFB, and 20% returned more than once after being hazed from EAFB. We identified three staging areas, geese utilized prior to entering EAFB, and three post-hazing dispersal sites. Collared geese began moving onto EAFB 30–40 days post-molt with the largest proportions moving onto EAFB 70–90 days post-molt. We observed 75 neckbanded geese on AIA from seven molting sites, and 23% returned more than once after being hazed from AIA. We observed 141 neckbanded geese on MFA from 14 molting sites, and 21% returned more than once after being hazed from MFA. Our data indicated that as long as local goose populations increase, large numbers of Anchorage area geese are likely to enter one of the airports creating a variety of management problems. Hazed geese returning to airports multiple times present a special hazard to aircraft safety because they appear to have become habituated to non-lethal scare tactics. We recommend an integrated management approach to limit the Anchorage area goose population utilizing various control techniques which are acceptable to Anchorage residents while continuing the hazing program at area airports. 相似文献
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Summary In a cell-free homogenate of the blue-green algaMicrocystis aeruginosa gas vacuoles are assembled within the first half hour after incubation. 相似文献
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