渔业科学进展
漁業科學進展
어업과학진전
Progress In Fishery Sciences
2015年
5期
71-80
,共10页
陈大菾%黄倢%王海亮%李晋
陳大菾%黃倢%王海亮%李晉
진대첨%황첩%왕해량%리진
铵盐%乙醇%常温%RNA
銨鹽%乙醇%常溫%RNA
안염%을순%상온%RNA
Ammonium salt%Ethanol%Normal temperature%RNA
以不同浓度乙醇为溶剂,配制不同pH的硫酸铵和乙酸铵饱和溶液,测试其对凡纳滨对虾组织的渗透效应和在常温条件下对RNA的保护作用。因为乙酸铵在乙醇中的溶解度远高于硫酸铵,且其铵离子的摩尔浓度在0?80%的乙醇溶剂中比较稳定。研究结果显示,乙酸铵比硫酸铵能更快渗透到整体的对虾组织中,并达到更高的浓度。50%、70%及以上的乙醇为溶剂的饱和硫酸铵、50%乙醇为溶剂的饱和乙酸铵(pH=6.0)溶液保存RNA的完整性较好,RNA拷贝数较大,半衰期较长。综合分析,推荐50%乙醇的饱和乙酸铵溶液(pH=6.0)作为对虾组织样品的常温保存液,该结果为野外样品采集及常温运输提供了一种有效方法。
以不同濃度乙醇為溶劑,配製不同pH的硫痠銨和乙痠銨飽和溶液,測試其對凡納濱對蝦組織的滲透效應和在常溫條件下對RNA的保護作用。因為乙痠銨在乙醇中的溶解度遠高于硫痠銨,且其銨離子的摩爾濃度在0?80%的乙醇溶劑中比較穩定。研究結果顯示,乙痠銨比硫痠銨能更快滲透到整體的對蝦組織中,併達到更高的濃度。50%、70%及以上的乙醇為溶劑的飽和硫痠銨、50%乙醇為溶劑的飽和乙痠銨(pH=6.0)溶液保存RNA的完整性較好,RNA拷貝數較大,半衰期較長。綜閤分析,推薦50%乙醇的飽和乙痠銨溶液(pH=6.0)作為對蝦組織樣品的常溫保存液,該結果為野外樣品採集及常溫運輸提供瞭一種有效方法。
이불동농도을순위용제,배제불동pH적류산안화을산안포화용액,측시기대범납빈대하조직적삼투효응화재상온조건하대RNA적보호작용。인위을산안재을순중적용해도원고우류산안,차기안리자적마이농도재0?80%적을순용제중비교은정。연구결과현시,을산안비류산안능경쾌삼투도정체적대하조직중,병체도경고적농도。50%、70%급이상적을순위용제적포화류산안、50%을순위용제적포화을산안(pH=6.0)용액보존RNA적완정성교호,RNA고패수교대,반쇠기교장。종합분석,추천50%을순적포화을산안용액(pH=6.0)작위대하조직양품적상온보존액,해결과위야외양품채집급상온운수제공료일충유효방법。
Molecular biology technologies have been widely used in aquatic animal disease diagnosis and molecular pathology studies, which require high quality of nucleic acid. However, how to preserve sample quality at normal temperature is rarely investigated. In this study, we compared the preserving effect of RNA in the tissues of shrimpLitopenaeus vannamei stored in different preservation solutions at normal temperature to find one preservation solution that can be used at normal temperature and long-distance transportation. Saturated solutions of ammonium sulfate and ammonium acetate in a serial of solvents with different ethanol concentrations were used to preserve shrimp tissues. The results showed that ammonium acetate has higher solubility in ethanol solution than ammonium sulfate. The molarity of ammonium ion of ammonium acetate is steadier in 0?80% ethanol. Ammonium acetate penetrates into the tissue of intact shrimp faster and reaches a higher concentration than ammonium sulfate does. RNA with better integrity, higher copies, and longer half-life period was achieved in the tissue preserved in more than 70% ethanol, saturated ammonium sulfate in the solvent of 50% and 70% ethanol, saturated ammonium acetate in the solvent of 50% ethanol. These results suggest that the saturated ammonium acetate in 50% ethanol as the solvent (pH=6.0) can be used as sample preserving solution to preserve RNA in shrimp tissues at normal temperature.