{"id":477,"date":"2013-06-10T18:32:13","date_gmt":"2013-06-10T18:32:13","guid":{"rendered":"https:\/\/wordpress.clarku.edu\/debrobertson\/?page_id=477"},"modified":"2018-01-25T18:42:54","modified_gmt":"2018-01-25T18:42:54","slug":"f2-culture-media","status":"publish","type":"page","link":"https:\/\/wordpress.clarku.edu\/debrobertson\/laboratory-protocols\/f2-culture-media\/","title":{"rendered":"F\/2 Culture Media"},"content":{"rendered":"

F\/2 Medium Stock Solutions:<\/b><\/p>\n\n\n\n\n\n\n\n\n
\n

Quantity per L of Media<\/b><\/p>\n<\/td>\n

\n

Compound<\/p>\n<\/td>\n

\n

Stock Concentration<\/b><\/p>\n<\/td>\n

 <\/td>\n<\/tr>\n
1.0 ml<\/td>\nNaNO3<\/sub><\/td>\n75.0 g\/L dH2<\/sub>O<\/td>\nAutoclave separately<\/td>\n<\/tr>\n
1.0 ml<\/td>\nNaH2<\/sub>PO4<\/sub>.H2O<\/td>\n5.0 g\/L dH2<\/sub>O<\/td>\nAutoclave separately<\/td>\n<\/tr>\n
1.0 ml<\/td>\nNa2<\/sub>SiO3<\/sub>.9H2O<\/td>\n30.0 g\/L dH2<\/sub>O<\/td>\nAutoclave separately<\/td>\n<\/tr>\n
1.0 ml<\/td>\nf\/2 Trace Metal Solution<\/td>\n(see recipe below)<\/td>\nFilter sterilize<\/td>\n<\/tr>\n
0.5 ml<\/td>\nf\/2 Vitamin Solution<\/td>\n(see recipe below)<\/td>\nFilter sterilize<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Stock solutions (except vitamins) are considered 1000x. Seawater should be autoclaved and cooled prior to mixing the media.  We tend to sterile-filter artificial seawater rather than autoclaving to avoid reductions in dissolved carbon dioxide. A<\/p>\n

Add all the f\/2 components aseptically to your media once it has cooled to room temperature. For other algal groups (e.g. dinoflagellates, coccolithophores) use f\/2-Si medium (prepare as for f\/2 medium but omit Na2<\/sub>SiO3<\/sub>.9H2O). Sterile stock solutions can be stored at room temperature. Once opened, stock solutions should be stored at 4 \u00b0C.<\/p>\n

Trace Metal Solution<\/b><\/p>\n

To 950 mL distilled H2O add:<\/p>\n\n\n\n\n\n\n\n\n\n\n
\n

Quantity<\/b><\/p>\n<\/td>\n

\n

Compound<\/b><\/p>\n<\/td>\n

\n

Stock Solution<\/b><\/p>\n<\/td>\n<\/tr>\n

1.3 g<\/td>\nFeCl3<\/sub>.6H2<\/sub>O<\/td>\n <\/td>\n<\/tr>\n
8.7 g<\/td>\nNa2<\/sub>EDTA.2H2<\/sub>O<\/td>\n <\/td>\n<\/tr>\n
1.0 ml<\/td>\nCuSO4<\/sub>.5H2<\/sub>O<\/td>\n980 mg \/ 100 ml dH2<\/sub>0<\/td>\n<\/tr>\n
1.0 ml<\/td>\nNa2<\/sub>MoO4<\/sub>.2H2<\/sub>O<\/td>\n630 mg \/ 100 ml dH2<\/sub>0<\/td>\n<\/tr>\n
1.0 ml<\/td>\nZnSO4<\/sub>.7H2<\/sub>O<\/td>\n2.2 g \/ 100 ml dH2<\/sub>0<\/td>\n<\/tr>\n
1.0 ml<\/td>\nCoCl2<\/sub>.6H2<\/sub>O<\/td>\n1.0 g \/ 100 ml dH2<\/sub>0<\/td>\n<\/tr>\n
1.0 ml<\/td>\nMnCl2<\/sub>.4H2<\/sub>O<\/td>\n18.0 g \/ 100 ml dH2<\/sub>0<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Initially, the solution will be cloudy. Add 1N NaOH to adjust pH to about 4.5 to clear the solution. Bring final volume to one liter.<\/p>\n

Vitamin Solution<\/b><\/p>\n

To 950 mL dH2O add:<\/p>\n\n\n\n\n\n\n
\n

Quantity<\/b><\/p>\n<\/td>\n

\n

Compound<\/b><\/p>\n<\/td>\n

\n

1o<\/sup> Stock Solution<\/b><\/p>\n<\/td>\n

\n

Tip<\/b><\/p>\n<\/td>\n<\/tr>\n

1.0 ml<\/td>\nVitamin B12 <\/sub><\/td>\n10 mg \/ 10 ml dH2<\/sub>O<\/td>\n <\/td>\n<\/tr>\n
1.0 ml<\/td>\nBiotin<\/td>\n10 mg \/ 10 ml dH2<\/sub>O<\/td>\nDissolve in a little 0.1N NaOH first<\/td>\n<\/tr>\n
200.0 mg<\/td>\nThiamine HCl<\/td>\n <\/td>\n <\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Make final volume up to 1.0 L with dH2<\/sub>O. Store primary stock solutions in freezer. Store completed vitamin solution in the refrigerator or freeze small aliquots.<\/p>\n

 <\/b><\/p>\n

References<\/b><\/p>\n

Guillard R.R.L. and Ryther J.H. (1962) Studies of marine planktonic diatoms. I. Cyclotella nana Hustedt and Detonula confervacea Cleve. Can. J. Microbiol. 8: 229-239.<\/p>\n

Guillard R.R.L. (1975) Culture of phytoplankton for feeding marine invertebrates. in “Culture of Marine Invertebrate Animals.” (eds: Smith W.L. and Chanley M.H.) Plenum Press, New York, USA. pp 26-60.<\/p>\n

 <\/p>\n

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

F\/2 Medium Stock Solutions: Quantity per L of Media Compound Stock Concentration   1.0 ml NaNO3 75.0 g\/L dH2O Autoclave separately 1.0 ml NaH2PO4.H2O 5.0 g\/L dH2O Autoclave separately 1.0 ml Na2SiO3.9H2O 30.0 g\/L dH2O Autoclave separately 1.0 ml f\/2 …<\/p>\n

F\/2 Culture Media<\/span> Read More »<\/a><\/p>\n","protected":false},"author":72,"featured_media":0,"parent":8,"menu_order":0,"comment_status":"closed","ping_status":"open","template":"","meta":{"ngg_post_thumbnail":0,"footnotes":""},"class_list":["post-477","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"https:\/\/wordpress.clarku.edu\/debrobertson\/wp-json\/wp\/v2\/pages\/477","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/wordpress.clarku.edu\/debrobertson\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/wordpress.clarku.edu\/debrobertson\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/wordpress.clarku.edu\/debrobertson\/wp-json\/wp\/v2\/users\/72"}],"replies":[{"embeddable":true,"href":"https:\/\/wordpress.clarku.edu\/debrobertson\/wp-json\/wp\/v2\/comments?post=477"}],"version-history":[{"count":0,"href":"https:\/\/wordpress.clarku.edu\/debrobertson\/wp-json\/wp\/v2\/pages\/477\/revisions"}],"up":[{"embeddable":true,"href":"https:\/\/wordpress.clarku.edu\/debrobertson\/wp-json\/wp\/v2\/pages\/8"}],"wp:attachment":[{"href":"https:\/\/wordpress.clarku.edu\/debrobertson\/wp-json\/wp\/v2\/media?parent=477"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}