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Marine invertebrate sample (Sample ID: NIO_SA_An_15) was collected from deep sea (Anjuna Beach, Goa, India) in sterile polypropylene bag and stored at 4˚C. The invertebrate (~10 g) sample was washed thrice with sterile demineralized water to remove the adhering particles like sand, sea weeds and then crushed in the sterile mortar and pestle. The crushed sample was serially diluted to 10 −8 . Last three dilutions viz. 10 −6 , 10 −7 and 10 −8 were surface spreaded on the actinomycetes isolation agar (AIA) medium (Himedia, India) prepared in 75% (v/v) artificial sea water (AS-AIA). The plates were incubated at 30˚C till 30 days and observed periodically for the growth of actinomycetes colonies. The well isolated colonies were picked up, purified and grown on AS-AIA slants (made in 75% ASW). The well grown isolates on the slants were preserved at 4˚C. |
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| 海洋无脊椎动物样品（样品号NIO_SA_An_15）是使用无菌聚丙烯包采自深海（印度果阿安朱纳海滩），并于4˚C下保存。本样品（~10g）首先使用无菌脱盐水洗涤3次以除去粘附的粒子（如沙粒）以及海草，然后使用无菌研钵粉碎。粉碎的样品使用无菌水稀释至10−8。最后三种（梯度）稀释液，即10−6、10−7和10−8，在使用75% (v/v)的人造海水配置的AIA培养基（AS-AIA）表面散开。再于30˚C下孵育30天，并定期观察放线菌菌落的生长。取出得到良好分离的菌落，纯化，并在AS-AIA斜面培养基上生长。生长良好的分离物在4˚C下保存。 |
2楼2014-10-25 13:54:35
