{"id":2546,"date":"2014-04-25T09:34:47","date_gmt":"2014-04-25T01:34:47","guid":{"rendered":"http:\/\/www.leadingtec.cn\/?p=2546"},"modified":"2014-04-25T13:31:15","modified_gmt":"2014-04-25T05:31:15","slug":"esaw-culture-medium","status":"publish","type":"post","link":"https:\/\/www.leadingtec.cn\/esaw-culture-medium.html","title":{"rendered":"ESAW\u57f9\u517b\u6db2\uff08ESAW\/HESNW Culture Medium\uff09"},"content":{"rendered":"

ESAW\u57f9\u517b\u6db2\u57f9\u517b\uff0c\u6587\u4ef6\u8d44\u6599\u6765\u6e90\u4e8ebotany.ubc.ca<\/p>\n

The medium given below can be made with either artificial seawater or natural seawater.<\/p>\n


\n<\/span>Artificial Seawater<\/b><\/span><\/p>\n

Make two separate solutions: Solution I and Solution II. Add salts in order to distilled
\ndeionized water (DDW). When the salts in each are completely dissolved mix
\nsolutions together. Dilute to total volume with DDW.<\/span><\/p>\n

 <\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
Table 1. ESAW Salts<\/span><\/b><\/caption>\n
Solution I:
\nAnhydrous salts<\/b><\/span><\/td>\n		Molecular Weight
\n(g\/mol)<\/b><\/span><\/td>\n		Amount to weigh:
\n(g\/l solution)<\/b><\/span><\/td>\n		Concentration
\n(mM)<\/b><\/span><\/td>\n<\/tr>\n
NaCl<\/span><\/td>\n58.44<\/span><\/td>\n20.756<\/span><\/td>\n362.7<\/span><\/td>\n<\/tr>\n
Na2<\/sub>SO4<\/sub><\/span><\/td>\n142.04<\/span><\/td>\n3.477<\/span><\/td>\n25.0<\/span><\/td>\n<\/tr>\n
KCl<\/span><\/td>\n74.56<\/span><\/td>\n0.587<\/span><\/td>\n8.03<\/span><\/td>\n<\/tr>\n
NaHCO3<\/sub><\/span><\/td>\n84<\/span><\/td>\n0.17<\/span><\/td>\n2.067<\/span><\/td>\n<\/tr>\n
KBr<\/span><\/td>\n119.01<\/span><\/td>\n0.0845<\/span><\/td>\n0.725<\/span><\/td>\n<\/tr>\n
H3<\/sub>BO3<\/sub><\/span><\/td>\n61.83<\/span><\/td>\n0.022<\/span><\/td>\n0.372<\/span><\/td>\n<\/tr>\n
NaF<\/span><\/td>\n41.99<\/span><\/td>\n0.0027<\/span><\/td>\n0.0657<\/span><\/td>\n<\/tr>\n
Solution II – Hydrated salts<\/span><\/b><\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n
MgCl2<\/sub>\u00a0.<\/sup>\u00a06H2<\/sub>O<\/span><\/td>\n203.33<\/span><\/td>\n9.395<\/span><\/td>\n47.18<\/span><\/td>\n<\/tr>\n
CaCl2<\/sub>\u00a0.<\/sup>\u00a02H2<\/sub>O<\/span><\/td>\n147.03<\/span><\/td>\n1.316<\/span><\/td>\n9.134<\/span><\/td>\n<\/tr>\n
SrCl2<\/sub>\u00a0.<\/sup>\u00a06H2<\/sub>O<\/span><\/td>\n266.64<\/span><\/td>\n0.0214<\/span><\/td>\n0.082<\/span><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
Table 2. Nutrient Enrichment Stocks<\/b><\/span><\/caption>\n
<\/td>\nEnrichment Stocks<\/span><\/td>\nStock Conc.(g\/L)<\/span><\/td>\nFinal Conc.(\u00b5M)<\/span><\/td>\n<\/tr>\n
1<\/span><\/td>\nNaNO3<\/sub><\/span><\/td>\n46.67<\/span><\/td>\n549.1<\/span><\/td>\n<\/tr>\n
2\u00a0<\/span>*<\/span><\/td>\nNa2<\/sub>glycerophosphate<\/span><\/td>\n6.67<\/span><\/td>\n21.8<\/span><\/td>\n<\/tr>\n
3<\/span><\/td>\nNa2<\/sub>SiO3\u00a0<\/sub>.<\/sup><\/b>\u00a09H2<\/sub>O<\/span><\/td>\n15.00<\/span><\/td>\n105.6<\/span><\/td>\n<\/tr>\n
4\u00a0<\/span>**<\/span><\/td>\nNa2<\/sub>EDTA\u00a0.<\/sup><\/b>\u00a02H2<\/sub>O<\/span><\/td>\n3.64<\/span><\/td>\n9.81<\/span><\/td>\n<\/tr>\n
***<\/span><\/td>\nFe(NH4<\/sub>)2<\/sub>(SO4<\/sub>)2\u00a0<\/sub>.<\/sup><\/b>\u00a06H2<\/sub>O<\/span><\/td>\n2.34<\/span><\/td>\n5.97<\/span><\/td>\n<\/tr>\n
<\/td>\nFeCl3\u00a0<\/sub>.<\/sup><\/b>\u00a06H2<\/sub>O<\/span><\/td>\n0.16<\/span><\/td>\n0.592<\/span><\/sup><\/td>\n<\/tr>\n
5<\/span><\/td>\nMnSO4\u00a0<\/sub>.<\/sup><\/b>\u00a04H2<\/sub>O<\/span><\/td>\n0.54<\/span><\/td>\n2.42<\/span><\/td>\n<\/tr>\n
<\/td>\nZnSO4\u00a0<\/sub>.<\/sup><\/b>\u00a07H2<\/sub>O<\/span><\/td>\n0.073<\/span><\/td>\n0.254<\/span><\/td>\n<\/tr>\n
<\/td>\nCoSO4\u00a0<\/sub>.<\/sup><\/b>\u00a07H2<\/sub>O<\/span><\/td>\n0.016<\/span><\/td>\n0.0569<\/span><\/td>\n<\/tr>\n
<\/td>\nNa2<\/sub>MoO4\u00a0<\/sub>.<\/sup><\/b>\u00a02H4<\/sub>O<\/span><\/td>\n0.126<\/span><\/td>\n0.520<\/span><\/td>\n<\/tr>\n
<\/td>\n**<\/span>\u00a0Na2<\/sub>EDTA\u00a0.\u00a0<\/sup><\/b>2H2<\/sub>O<\/span><\/td>\n1.89<\/span><\/td>\n5.05<\/span><\/td>\n<\/tr>\n
6<\/span><\/td>\nH3<\/sub>BO3<\/sub><\/span><\/td>\n3.80<\/span><\/td>\n61.46<\/span><\/td>\n<\/tr>\n
7<\/span><\/td>\nNa2<\/sub>SeO3<\/sub><\/span><\/td>\n0.00173<\/span><\/td>\n0.001<\/span><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n


\nEnrichment stocks can be refrigerated. Adjust Solution #5 to pH 6 using solid NaCO3<\/sub>\u00a0(about 2 g). Solution # 4 can be heated to dissolve the iron.<\/span><\/p>\n

Notes:<\/span><\/p>\n

*<\/span>\u00a0Can be replaced with an equimolar stock of Na2<\/sub>HPO4<\/sub>
\n<\/span>**<\/span>\u00a0Add before the trace metals.
\n<\/span>***<\/span>\u00a0Can be replaced with an equimolar stock of FeCl3<\/sub><\/span><\/p>\n

 <\/p>\n\n\n\n\n\n\n
Table 3. Vitamin Stocks<\/b><\/span><\/caption>\n
Vitamin Stock<\/span><\/td>\nStock Conc.(g\/L)<\/span><\/td>\nFinal Conc.(mM)<\/span><\/td>\n<\/tr>\n
Thiamine<\/span><\/td>\n0.1<\/span><\/td>\n2.97 x 10-1<\/sup><\/span><\/td>\n<\/tr>\n
Vitamin B12<\/span><\/td>\n0.002<\/span><\/td>\n1.47 x 10-3<\/sup><\/span><\/td>\n<\/tr>\n
Biotin<\/span><\/td>\n0.001<\/span><\/td>\n4.09 x 10-3<\/sup><\/span><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n


\nVitamin stocks should be stored frozen.<\/span><\/p>\n

TO MAKE 1 LITRE OF HESNW<\/b><\/span><\/p>\n

1) Filter natural seawater through 0.45 \u00b5m membrane filter with a glass fiber prefilter.<\/p>\n

2) Use a flask that has been acid-washed in 10% HCl and rinsed in distilled water before first use.<\/p>\n

3) Pour 1 L filtered seawater or artificial seawater into flask. Add the following while mixing with a stir-bar:<\/p>\n

a) 6 ml Combined Stock (Enrichment stocks #1,2,4-7)
\n(OR 1 ml of each individual stock solution)
\nb) 2 ml Silica Stock (Enrichment stock #3)
\nc) 2 ml Vitamin Stock<\/p>\n

4) To reduce precipitation during autoclaving add:<\/p>\n

d) 1.44 ml of 1N HCl
\ne) 0.12 g Sodium bicarbonate<\/p>\n

5) Autoclave 20 minutes for one litre, ~ 30 minutes for 2 litres. Autoclaving for longer periods will result in more precipitation. Flasks should not be filled up more than 2\/3 of their capacity, to avoid boiling over during sterilization.<\/p>\n

6) Let stand for 2 days to allow pH to equilibrate (should be ~ pH 8.2).<\/p>\n

7) Filter aseptically through a glass fiber filter to remove precipitate. Some taxa appear to be sensitive to it (optional).<\/p>\n

Reference<\/b><\/span><\/p>\n

Harrison, P.J., R.E. Waters and F.J.R. Taylor. 1980. A broad spectrum artificial medium for coastal and open ocean phytoplankton. J. Phycol. 16:28-35.<\/p>\n

 <\/p>\n","protected":false},"excerpt":{"rendered":"

ESAW\u57f9\u517b\u6db2\u57f9\u517b\uff0c\u6587\u4ef6\u8d44\u6599\u6765\u6e90\u4e8ebotany.ubc.ca<\/p>\n","protected":false},"author":2,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[100],"tags":[229],"class_list":["post-2546","post","type-post","status-publish","format-standard","hentry","category-medium","tag-229"],"aioseo_notices":[],"_links":{"self":[{"href":"https:\/\/www.leadingtec.cn\/wp-json\/wp\/v2\/posts\/2546"}],"collection":[{"href":"https:\/\/www.leadingtec.cn\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.leadingtec.cn\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.leadingtec.cn\/wp-json\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/www.leadingtec.cn\/wp-json\/wp\/v2\/comments?post=2546"}],"version-history":[{"count":0,"href":"https:\/\/www.leadingtec.cn\/wp-json\/wp\/v2\/posts\/2546\/revisions"}],"wp:attachment":[{"href":"https:\/\/www.leadingtec.cn\/wp-json\/wp\/v2\/media?parent=2546"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.leadingtec.cn\/wp-json\/wp\/v2\/categories?post=2546"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.leadingtec.cn\/wp-json\/wp\/v2\/tags?post=2546"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}