Collections of the Biological Culture Service - IBVF

i 1. CYANOBACTERIAS

Cyanobacteria are microorganisms that, in a prokaryotic organization, are capable of carrying out oxygenic photosynthesis similar to that of higher plants. They are grouped into five Sections, whose basic characteristics are set out below. The Biological Cultures Service of the Isla de la Cartuja Scientific Research Centre has a collection of cyanobacteria with representatives for each of the Sections.

Growing conditions

The most commonly used media for the cultivation of cyanobacteria in the collection are BG11 medium ₀ and ASN-III (RippKa et al, 1979). For solid medium it is used Bacto-Agar (1% p/v).

The different strains are cultivated under photoautotrophic conditions in a thermostatically controlled chamber at 30°C.

Sections

Unicellular; single cells or forming aggregates surrounded by additional outer cell wall layers	Reproduction by binary division or by budding		Section I
	Reproduction by multiple fission giving rise to small daughter cells (baeocytes) or by multiple fission and binary fission.		Section II
Filamentous; a trichome (chain of cells) that grows by division of intercalary cells	Reproduction by formation of hormogonia, by random breaking of the trichome, and (4th and 5th s.) sometimes by germination of akinetes.	Trichome always composed of vegetative cells	Division in a plane Section III
		In the absence of combined nitrogen the trichomes have heterocysts, some also akinetes	Division in a plane Section IV
			Division into more than one plane Section V

List of strains (Photos of the collection-pdf)

NAME	Section	Growing conditions
Acaryochloris marina NBRC 102967 (MBIC11017)	I	Medio Acaryochloris (medio 963, NITE)
Anabaena azollae AS-DS	IV	BG11₀ +/- fuente de N (nitrato o amonio).
Anabaena cylindrica ATCC 29414	IV	BG11₀ +/- fuente de N (nitrato o amonio).
Anabaena sp. ATCC 33047	IV	BG11₀ +/- fuente de N (nitrato o amonio).
Anabaena sp. PCC 6411 (ATCC 27898)	IV	BG11₀ +/- fuente de N. No fija N2 en aerobiosis.
Anabaena sp. PCC 7119 (ATCC 29151)	IV	BG11₀ +/- fuente de N. (nitrato o amonio).
Anabaena sp. PCC 7120 (ATCC 27893)	IV	BG11₀ +/- fuente de N (nitrato, amonio o urea).
Anabaena variabilis ATCC 29413 (PCC 7937)	IV	BG11₀ +/- fuente de N (nitrato o amonio).
Anabaena sp. Universidad de Leningrado 458 (PCC 7118)	IV	BG11₀ +/- fuente de N (nitrato o amonio). No fija N2 en aerobiosis.
Anabaena sp. Universidad de Tokio M131	IV	BG11₀ +/- fuente de N (nitrato o amonio). No fija N2 en aerobiosis.
Anabaena variabilis ATCC 29413 – FD	IV	BG11₀ +/- fuente de N (nitrato o amonio).
Anabaena variabilis ATCC 29413 – P9	IV	BG11₀ +/- fuente de N (nitrato o amonio).
Anacystis nidulans L-1402-1	I	BG11.
Arthrospira maxima	III	BG11C.
Calothrix (Tolypothrix) sp. PCC 7601 (Fremyella diplosiphon UTEX 481)	IV	BG11₀ +/- fuente de N (nitrato o amonio). Fija dinitrógeno en anaerobiosis.
Calothrix sp. PCC 9327	IV	BG11₀ +/- fuente de N (nitrato o amonio).
Chlorogloeopsis sp. PCC 6912 (ATCC 29193)	V	BG11₀ +/- fuente de N (nitrato o amonio).
Chroococcidiopsis thermalis PCC 7203 (Myxosarcina chroococcoides)	II	BG11₀ +/- fuente de N (nitrato o amonio). Fija dinitrógeno en anaerobiosis
Chroococcidiopsis sp. PCC 9317	II	BG11
Cyanothece sp. PCC 7425	I	BG11₀ +/- fuente de N (nitrato o amonio). Fija dinitrógeno en anaerobiosis
Cyanothece sp. PCC 9318	I	BG11
Dermocarpa sp. PCC 7437 (Stanieria) (ATCC 29371)	II	BG11
Fischerella muscicola PCC 73103	V	BG11₀ +/- fuente de N (nitrato o amonio).
Fischerella muscicola PCC 7414	V	BG11₀ +/- fuente de N (nitrato o amonio).
Fischerella muscicola UTEX 1829	V	BG11₀ +/- fuente de N (nitrato o amonio).
Gloeobacter violaceus sp. PCC 7421	I	BG11
Gloeocapsa sp. PCC 9319	I	BG11
Geitlerinema sp. PCC 9326	III	BG11
Leptolyngbya sp. PCC 9324	III	BG11
Lyngbya sp. PCC 8106	I	BG11/ASN-III (1:1). Fija dinitrógeno en anaerobiosis.
Nodularia chucula	IV	BG11₀ +/- fuente de N (nitrato o amonio)
Nostoc ellipsosporum (Univ. de Göttingen B1453-79)	IV	BG11₀ +/- fuente de N (nitrato o amonio).
Nostoc paludosum PCC 9206	IV	BG11₀ +/- fuente de N (nitrato o amonio).
Nostoc sp. ATCC 29133 (PCC 73102) N. punctiforme	IV	BG11₀ + TES 10 mM pH 7,5 +/- fuente de N (nitrato o amonio)
Nostoc sp. 78-IIA-E (ATCC 43237)	IV	BG11₀ +/- fuente de N (nitrato o amonio).
Nostoc sp. caquena	IV	BG11₀ +/- fuente de N (nitrato o amonio)
Nostoc sp. llaita	IV	BG11₀ +/- fuente de N (nitrato o amonio).
Nostoc sp. PCC 6705 (ATCC 29131)	IV	BG11₀ +/- fuente de N (nitrato o amonio).
Nostoc sp. PCC 6719 (ATCC 29105)	IV	BG11₀ +/- fuente de N (nitrato o amonio).
Nostoc sp. PCC 6720 (ATCC 27899)	IV	BG11₀ +/- fuente de N (nitrato o amonio).
Nostoc sp. PCC 7107 (ATCC 29150)	IV	BG11₀ +/- fuente de N (nitrato o amonio).
Nostoc sp. PCC 7413 (ATCC 29106)	IV	BG11₀ +/- fuente de N (nitrato o amonio).
Nostoc sp. PCC 9201	IV	BG11₀ +/- fuente de N (nitrato o amonio)
Nostoc sp. PCC 9202	IV	BG11₀ +/- fuente de N (nitrato o amonio).
Nostocsp. PCC 9203	IV	BG11₀ +/- fuente de N (nitrato o amonio)
Nostoc sp. PCC 9204	IV	BG11₀ +/- fuente de N (nitrato o amonio)
Nostoc sp. PCC 9329	IV	BG11₀ +/- fuente de N (nitrato o amonio).
Nostoc sp. PCC 9331	IV	BG11₀ +/- fuente de N (nitrato o amonio).
Oscillatoria sp. PCC 9325	III	BG11
Phormidium laminosum (Agardh) Gom. OH-1-pC11	III	BG11. Termófilo (45°C)
Phormidium persicinum 80.79	III	Medio f/2 de Guillard
Pseudanabaena sp. PCC 6903 (ATCC 27190)	III	BG11
Pseudanabaena sp. PCC 9330	III	BG11
Scytonema sp. PCC 7110 (ATCC 29171)	IV	BG11₀ +/- fuente de N (nitrato o amonio).
Spirulina sp. PCC 6313 (ATCC 29542)	III	BG11/ASNIII (1:1)
Synechococcus A6	I	BG11.
Synechococcus M4-	I	BG11.
Synechococcus sp. PCC 6301 (ATCC 27144) “S. leopoliensis”	I	BG11
Synechococcus sp. PCC 7202 (ATCC 29140)	I	BG11 + 10% ASN-III.
Synechococcus sp. PCC 7335	I	ASN-III. Fija dinitrógeno en anaerobiosis.
Synechococcus sp. PCC 7942 (Anacystis nidulans R2)	I	BG11
Synechococcus sp. PCC 9321	I	BG11
Synechocystis sp. PCC 6308 (ATCC 27150)	I	BG11
Synechocystis sp. PCC 6803 (ATCC 27184)	I	BG11
Synechocystis sp. PCC 9320	I	BG11
Synechocystis sp. PCC 9322	I	BG11
Synechocystis sp. PCC 9323	I	BG11

2. ALGAE

Algae are eukaryotic organisms (with numerous exceptions), aquatic (with frequent exceptions), autotrophic with oxygenic photosynthesis and with a smaller size and a less complex structure than terrestrial plants. It is a non-monophyletic group of organisms for which, depending on the authors, different phyla or divisions can be distinguished: Glaucophyta, Euglenophyta, Cryptophyta, Haptophytes, Dinophytes, Ochrophytes, Rhodophytes and Chlorophytes (see main characteristics in the following table). The Biological Cultures Service of the Isla de la Cartuja Scientific Research Centre has a collection of microalgae with representatives of these Divisions.

Growing conditions

The medium used for the cultivation of the microalgae in the collection varies depending on the strain in question. As a general rule, the microalgae are maintained in a solidified medium with Bacto-agar (1% w/v), or in a liquid medium in culture tubes.

The different strains are cultivated under photoautotrophic conditions in a thermostatically controlled chamber at 25°C.

Phyla or divisions

	Photosynthetic pigments	Warehouse products	Cellular cover
Glaucofitas	Clorofila a, ficocianina, aloficocianina, ficoeritrina, b-caroteno, xantofilas	Almidón	Algunas celulósicas
Euglenofitas	Clorofila a y b, b-caroteno, otros carotenos y xantofilas	Paramilon	Película proteínica bajo membrana plasmática
Cryptofitas	Clorofila a y c ficocianina, aloficocianina, ficoeritrina, ay b-caroteno, xantofilas	Almidón	Película proteínica bajo la membrana plasmática
Haptofitas	Clorofila a y c, b-caroteno, xantofilas	Crisolaminarina	Escamas de CaCO₃
Dinofitas	Clorofila a y c, b-caroteno, xantofilas	Almidón	Placas celulósicas bajo la membrana plasmática
Ochrofitas (diatomeas, crisófitas, algas marrones…) (eustigmatofitas)	Clorofila a y c, b-caroteno, xantofilas Clorofila a, b-caroteno, xantofilas	Crisolaminarina, lípidos Crisolaminarina	Algunas desnudas, otras con escamas orgánicas/silíceas, otras con celulosa o alginato Presente, pero de composición desconocida
Rodofitas	Clorofila a ficocianina, aloficocianina, ficoeritrina, ay b-caroteno, xantofilas	Almidón florideano	Celulosa, polisacáridos sulfatados; algunos calcificados
Clorofitas	Clorofila a y b, b-caroteno, otros carotenos y xantofilas	Almidón	Pared de celulosa u otros polímeros; escamas sobre algunos; otros desnudos; otros calcificados

(Algae, 2000, Graham LE and Wilcox LW eds., Prentice Hall Inc, USA)

List of strains (photos of the collection-pdf)

NAME	Type	Growing conditions
Astasia longa 1204-17a	Euglenofita	Medio Euglena
Chlamydomonas augustae (C. nivalis o Chloromonas reticulata) CCAP11/51, SAG 26.86	Clorofita	SUEOKA. Nit+
Chlamydomonas euryale 25-89	Clorofita	SUEOKA. Nit –
Chlamydomonas reinhardtii 21gr (+)	Clorofita	SUEOKA. Nit+
Chlamydomonas reinhardtii 137c (+)	Clorofita	SUEOKA. Nit –
Chlamydomonas reinhardtii CC124 (-)	Clorofita	SUEOKA. Nit –
Chlamydomonas reinhardtii CC621 (-)	Clorofita	SUEOKA. Nit –
Chlamydomonas reinhardtii CF10 (-)	Clorofita	SUEOKA. Nit+
Chlamydomonas reinhardtii CF11 (-)	Clorofita	SUEOKA. Nit –
Chlamydomonas reinhardtii CF14 (+)	Clorofita	SUEOKA. Nit+
Chlamydomonas reinhardtii CW15-325 (+)	Clorofita	SUEOKA. Nit + Arg –
Chlamydomonas reinhardtii CW15 (-)	Clorofita	SUEOKA. Nit +
Chlamydomonas reinhardtii 704 (+)	Clorofita	SUEOKA. Nit +
Chlorella sp. CCAP 211/84	Clorofita	Medio H. Nit +
Chlorella fusca (C. emersonii) 211-8b	Clorofita	SUEOKA. Nit +
Chlorella salina 8-86	Clorofita	Medio f/2 de Guillard
Chlorella sorokiniana 211-32	Clorofita	SUEOKA. Nit +
Chlorella stigmatofora 9.86	Clorofita	Medio ASW
Chlorella vulgaris 9-88 (UAM 101)	Clorofita	SUEOKA. Nit +/-
Chlorella zofingiensis 211/14	Clorofita	SUEOKA. Nit +/-
Chlorococcum citriforme 62.80	Clorofita	SUEOKA. Nit +
Chromera velia CCAP 1602/1	Apicomplexa	Medio ASW
Coelastrum proboscideum (var. dilatatum) 217-2	Clorofita	SUEOKA. Nit +
Coelastrum proboscideum (var. gracile) 217-3	Clorofita	SUEOKA. Nit +
Cryptomonas sp. 06/0101	Criptofita	Medio ASW
Cyanidium caldarium 16-91	Rodofita	Medio Cyanidium
Cyanophora paradoxa	Glaucofita	BG11 (2 mM NaNO₃)
Dunaliella salina	Clorofita	Medio Dunaliella
Euglena gracilis SAG 5-9	Euglenofita	Medio H
Euglena gracilis SAG 5-15	Euglenofita	Medio H
Euglena gracilis SAG 5-25	Euglenofita	Medio H
Galdieria sulphuraria 74W	Rodofita	Medio Cyanidium
Haematococcus lacustris 192.80	Clorofita	Medio H
Haematococcus lacustris ATCC 30453	Clorofita	Medio H
Haematococcus pluvialis CCAP 34/8	Clorofita	Medio H
Haematococcus pluvialis SAG 192.80	Clorofita	Medio H
Haematococcus pluvialis SAG 34-1b	Clorofita	Medio H
Heterocapsa sp. 09/0601	Dinofita	Medio f/2 de Guillard
Isochrysis galbana 05/0401	Ochrofita	Medio ASW
Micromonas commoda RCC 827	Prasinofícea	Medio ASW
Monoraphidium braunii 202.7d	Clorofita	SUEOKA. Nit +
Monoraphidium circinale	Clorofita	Medio f/2 de Guillard (0,25x)
Muriellopsis sp. IBVF-L4	Clorofita	SUEOKA. Nit +
Nannochloropsis gaditana	Ochrofita	Medio ASW
Nannochloris atomus SAG 14.87	Clorofita	Medio ASW

Nannochloropsis oculata 38.85	Ochrofita	Medio ASW
Nannochloropsis salina 40.85	Ochrofita	Medio ASW
Navicula pelliculosa 1050-3	Ochrofita	Específico
Neochloris oleoabundans UTEX 1185	Clorofita	BG11
Neospongiococcum (Chlorococcum) gelatinosum 64.80	Clorofita	SUEOKA. Nit +
Nephroselmis olivacea CCAP 1960/4B	Prasinofícea	3N-BBM+V (CCAP)
Ochromonas danica 933-7	Ochrofita	Medio Euglena
Ostreococcus tauri RCC745	Clorofita	Medio ASW
Phaeodactylum tricornutum 07/0502	Ochrofita	Medio ASW
Phaeodactylum tricornutum CCAP 1055/1	Ochrofita	Medio ASW
Porphyridium purpureum 1380-1a	Rodofita	Medio ASW
Poterioochromonas malhamensis CCAP 933/1C	Ochrofita	Medio Euglena
Pycnococcussp. RCC245	Prasinofícea	Medio ASW
Pycnococcus provasolii CCAP 190/1	Prasinofícea	Medio ASW
Rhodomonas baltica 06/0301	Criptofita	Medio ASW
Rhodomonas salina	Criptofita	Medio ASW
Scenedesmus vacuolatus SAG 211-15	Clorofita	SUEOKA. Nit +
Spirogyra sp SAG 170.80	Zygnematofícea	BG11
Tetracystis sp.	Clorofita	SUEOKA. Nit +
Tetraselmis chuii 8-6	Prasinofícea	Medio f/2 de Guillard.
Tetraselmis chui RCC128	Prasinofícea	Medio ASW
Tetraselmis suecica 03/0203	Clorofita	Medio ASW

3. ESCHERICHIA COLI

Escherichia coli is an enterobacteria that is widely used in molecular biology work. It is a fast-growing bacterium whose genome sequence is known and which allows the introduction of exogenous DNA, either by transformation, transduction or electroporation. The Biological Culture Service has a collection of Escherichia coli strains that is used by IBVF members in recombinant DNA work.

List of strains

CEPA	Genotype
E. coliBL21 (DE3)	F^– ompT (lon) hsdS(r_B-m_B-, una E. coli B) con DE3.
E. coliBL21 (DE3) lacI^q	F´::Tn10 proA+B+ lacIq D(lacZ)M15/ ompT (lon) hsdS (r_B-m_B-, una E. coli B) con DE3.
E. coli BL21 (DE3) pLysS	F^– ompT hsdS_B (r_B^–m_B^–) gal dcm (DE3) pLysS (CmR)
E. coliBL21 (DE3)(pIZ227)	pIZ227 porta, junto con lacIq, un gen de una lisozima específica de la polimerasa T7. Cm^r.
E. coliBL21 CodonPlus(DE3)-RIL	E. coliB F^– ompT (lon) hsdS(r_B-m_B-) dcm⁺ Tet^r gallHte (argU ileY LeuW Cm^r) -argU, AGA/AGG; IleY, AUA; LeuW, CUA-
E. coli BL21 Rosetta(DE3)pLys	F^–ompThsdS_B(r_B^–m_B^–) gal dcm (DE3)pLysRARE (Cm^R) (tRNAs para AGG, AGA, AUA, CUA, CCC y GGA en pLys)
E. coli BL21-AI	F^– ompT hsdS_B (r_B^–m_B^–)gal dcm araB::T7RNAP-tetA lon (tet^R)
E. coli BL21-T1^R	F^– ompT hsdS_B(r_B^–m_B^–) gal dcm tonA
E. coli C41(DE3)	F^– ompT hsdS_B (r_B– m_B-)gal dcm (DE3)
E. coli C43(DE3)	F^– ompT hsdS_B (r_B– m_B-)gal dcm (DE3)
E. coli CC118 lambda pir	Delta(ara-leu) araD delta lacX74 galE galK phoA20 thi rpsE rpoB argE(am)recA (lambda pir)
E. coli DB3.1	F– gyrA462 endA1 (sr1-recA) mcrB mrr hsdS20(rB-, mB-) supE44 ara-14 galK2 lacY1 proA2 rpsL20(SmR) xyl-5 – leu mtl1 ‘gateway’
E. coli DH10B T1R	F^– mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara leu) 7697 galU galK rpsL nupG λ– tonA
E. coliDH5a	F^–supE44 hsdR17 (r_K-r_K+) recA1 girA96 (Nal^r) endA1 thi-1 relA1 D(lacZYA-argF) (Ø80lacZDM15) U169
E. coli GM119	F^– dam-3 dcm-6 metB1 galK2 galT22 lacY1 tsx78 supE44 (thi1 tomA31 mtl1)
E. coliHB101	F^–hsdS20(r_B-m_B-) leu supE44 ara14 galK2 lacY1 proA2 rpsL20 xyl-5 mtl-1 recA13 mcrB
E. coliJM109	F´ traD36 lacIq D(lacZ)M15 proA+B+/e14-(McrA-) D(lac-proAB) thi gyrA96(Nal^r) endA1 hsdR17 (r_K-m_K+) relA1 supE44 recA1
E. coliJM109(DE3)	endA1, recA1, gyrA96, hsdR17(rk-, mk+), relA1, supE44, lambda -, Delta(lac-proAB), [F’, traD36, proAB, lacIqZdeltaM15]/ DE3
E. coli K12	Silvestre
E. coli Lemo21(DE3)	fhuA2 [lon] ompT gal (DE3)[dcm] DhsdS/pLemo (Cm^R)
E. coli S17-1 Lambda pir	RecA thi pro hsdR-M+ RP4:2-Tc:Mu:Km::Tn7 (Tpr Smr) (lambda pir)
E. coli SG13009 (pREP4)	F^–thi lac mtl; pREP4(Km^R, lacI^q)
E. coliXL1-Blue	F´::Tn10 proA+B+ lacIq D(lacZ)M15/recA1 endA1 gyrA96(Nal^r) thi hsdR17 (r_K-m_K+) supE44 relA1 lac
E.coli Solu (BL21)	F^– ompT hsdSB (r_B-m_B-) gal dcm (DE3)
E. coli XL1-Blue MRA (P2)	D(mcrA)183 D(mcrCB-hsdSMR-mrr)173 endA 1supE44 thi-1 gyrA96 relA1 lac (P2 lysogen)
E.coliMC1061	F^– araD139 D(ara-leu)7696 galE15 galK16 D(lac)X74 rpsL(Str^r) hsdR2 (r_K– m_K+) mcrA mcrB1

Culture medium BG11₀(Rippka et al, 1979)

Na₂CO₃	0,2 mM	H₃BO₃	46 µM
MgSO₄	0,3 mM	MnCl₂	9,1 µM
CaCl₂	0,24 mM	Na₂MoO₄	1,6 µM
K₂HPO₄	0,2 mM	ZnSO₄	0,8 µM
ácido cítrico	28,5 µM	CuSO₄	0,3 µM
citrato férrico-amónico (17% Fe)	6 mgl^-1	CoCl₂	0,2 µM
Na₂-EDTA	2,4 µM

The medium is prepared from a 100x concentrate lacking K2HPO4 and the nitrogen source, which are added before autoclaving.

For solid medium, Bacto-Agar is used at a final concentration of 1% w/v. This is autoclaved, in half the final volume, separately from the nutrients and mixed with them before pouring it into the Petri dishes.

Medium BG11

Medium BG11₀ supplemented with NaNO3 to a final concentration of 17.6 mM.

Medium BG11C BG11₀ medium supplemented with NaNO₃ at a final concentration of 17.6 mM and NaHCO₃ at a final concentration of 10 mM.edio BG11₀

Medium ASN-III (Rippka et al, 1979)

NaCl 25 g/l	citrato férrico-amónico (17% Fe) 0,003 g/l
MgSO₄ 3,5 g/l	Na₂-EDTA 0,0005 g/l
MgCl₂.6H₂O 0,24 mM	Na₂CO₃ 0,04 g/l
KCl 0,5 g/l	Metales traza (A5+Co) 1 ml/l
K₂HPO₄ 0,02 g/l
NaNO₃ 0,75 g/l
CaCl₂.2H₂O 0,5 g/l
ácido cítrico 0,003 g/l

Trace metals (A5 + Co)

H₃BO₃ 3,09 g/l

MnSO₄.4H₂O 2,9 g/l

ZnSO₄.7H₂O 0,29 g/l

(NH₄)₆Mo₇O₂₄^.4H₂O 0,62 g/l

CuSO₄^.5H₂O 124 mg/l

CoCl₂^.6H₂O 240 mg/l

Medium Sueoka

KH₂PO₄ 8,3 mM

K₂HPO₄ 5,3 mM

MgSO₄^.7H₂O 0,25 mM

CaCl₂^.2H₂O 0,133 mM

TRAZAS HUTNER 1x

NH₄Cl 9,35 mM o NaNO₃ 17,6 mM

For solid media, Bacto-Agar is used at a final concentration of 1% w/v. This is autoclaved, in half the final volume, separately from the nutrients and mixed with them before pouring it into the Petri dishes.

Traces Hutner 200x:

12,7 g of EDTA-Na^.2H₂O are dissolved in 250 ml of water.
They are dissolved 1 by 1 in this order, previously heating to 100ºC, in 500 ml of water:

– H₃BO₃ 2,28 g

– ZnSO₄^.7H₂O 4,40 g

– MnCl₂^.4H₂O 1,02 g

– FeSO₄^.7H₂O 1,00 g

– CoCl₂^.6H₂O 0,32 g

– CuSO₄^.5H₂O 0,32 g

– (NH₄)₆MoO₂₄^.4H₂O 0,22 g

The two previous solutions are mixed.
It is brought up to 100ºC and allowed to cool to 80-90ºC.
Adjust the pH to 6.8 with 20% KOH (temperature always >70ºC)
The volume is brought up to 1 litre with distilled water and stored in the dark for 2 days, after which it can be used.

Medium H

Medium Sueoka supplemented with 1.5 g/l of peptone proteose.

Medium Cyanidium

K₂HPO₄	0,3 g
CaCl₂	0,02 g
MgSO₄^.7H₂O	0,3 g
KNO₃	3,0 g
FeSO₄^.7H₂O	0,003 g
H₂SO₄	1,8 g
Metales traza (A5+Co)	1 ml/l
Agua (hasta 1 litro)

Trace metals (A5 + Co)

H₃BO₃ 3,09 g/l

MnSO₄.4H₂O 2,9 g/l

ZnSO₄.7H₂O 0,29 g/l

(NH₄)₆Mo₇O₂₄^.4H₂O 0,62 g/l

CuSO₄^.5H₂O 124 mg/l

CoCl₂^.6H₂O 240 mg/l

Medium Euglena

Acetato sódico 1 g

Extracto de carne 1 g

Bacto-triptona 2 g

Extracto de levadura 2 g

CaCl₂^.2H₂O 10 mg

Agua (hasta un litro)

For solid medium, Bacto-Agar is used at a final concentration of 1% w/v.

Medium f/2 of Guillard

Add 1 ml of stocks 1 and 3 to 998 ml of natural seawater.

Autoclave and add, once cold, 200 µl of stock 2 (sterilized by 0.2 µm filtration).

Stock 1. Trace metals.	Stock 2. Vitamin complex.	Stock 3.
EDTA-Na₂ 4,16 g	A. nicotínico 1 mg/ml	KNO₃ 15 g
FeCl₃^.6H₂O 3,15 g	A. para-aminobenzoico 1 mg/ml	NaH₂PO₄ 1,13 g
CuSO₄^.5H₂O 0,01 g	NaHCO3 0,5 mg/ml	Agua (hasta 100 ml)
ZnSO₄^.7H₂O 0,022 g	Tiamina HCl 0,5 mg/ml
CoCl₂^.6H₂O 0,01 g	Biotina 0,1 mg/ml
MnCl₂^.4H₂O 0,18 g	Vit. B12 0,035 mg/ml
Na₂MoO₄^.2H₂O 0,006g
Agua (hasta un litro)

MEDIUM ASW

Prepare a mixture of dry components composed of:

– NaCl 400 mM

-MgSO₄.7H₂O 20 mM

– CaCl₂.2H₂O 10 mM

– NaBr 1.7 mM

– KCl 10 mM

-MgCl₂ 20 mM

Sterilize by autoclave. Add to the previous mixture in sterility (per liter):

– H₃BO₃ 0.2 mM 1 ml

– Trizma 5%, pH 7.8 5 ml

– Stock 1 de f/2 1 ml

– Stock 3 de f/2 2 ml

– NaSiO₃.9H₂O 100 mM 1 ml

-Stock vitaminas: 0.5 ml

– Tiamina 0.2 mg/ml

– Biotina0.3 ug/ml .

-Vit. B12 1 ug/ml

Medium Navicula

NaNO₃1,18 mM

K₂HPO₄ 77 mM

Na₂SiO₃ 0,86 mM

Triptona 1 g/l

Concentrado BG11 2x

For solid media, Bacto-Agar is used at a final concentration of 1.5% w/v. This is autoclaved, in half the final volume, separately from the nutrients and mixed with them before pouring it into the Petri dishes.

Medium Acaryochloris

NaNO₃	200 mg
Na₂HPO₄	1,4 mg
K₂HPO₄	5 mg
NH₄Cl	2,68 mg
Fe-EDTA	5,2 mg
Mn-EDTA	0,332 mg
Na₂-EDTA	37,2 mg
ZnSO₄.7H₂O	0,023 mg
CoSO₄.7H₂O	0,014 mg
Na₂MoO₄.H₂O	0,0073 mg
CuSO₄.5H₂O	0,0025 mg
H₂SeO₃	0,0017 mg
MnCl₂.4H₂O	0,18 mg
Thiamin-HCl	0,2 mg
Biotin	0,0015 mg
VitaminB12	0,0015 mg
Sea water	hasta 1000 ml