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1. CYANOBACTERIA

INTRODUCTION

Cyanobacteria are microorganisms with a prokaryotic organization that are able to carry out an oxygenic photosynthesis similar to that of higher plants. They are grouped in five sections whose basic characteristics are exposed below. The CBCS cyanobacteria collection contains members of the different sections.

GROWTH CONDITIONS

Culture media most used for cyanobacteria growth are BG110 and ASN-III media (Rippka et al, 1979). Cyanobacteria are grown in phototrophy conditions at 25°C and low light intensity.

SECTIONS

Unicellular; cells single of forming aggregates held together by additional outer cell wall layers.

Reproduction by binary fission or by budding.

Section I

Reproduction by multiple fission giving rise to small daughter cells (baeocytes), or by both multiple fission and binary fission.

Section II

Filamentous; a trichome (chain of cells) which grows by intercalary cell division

Reproduction by random trichome breakage, by formation of hormogonia and (Sections IV and V only) sometimes by germination of akinetes

Trichome always composed only of vegetative cells

Division in only one plane

Section III

In the absence of combined nitrogen, trichome contains heterocysts; some also produce akinetes

Division in only one plane

Sección IV

Division in more than one plane

Section V

(Rippka et al, (1979) J. Gen. Microbiol. 111:1-61)

LIST OF CYANOBACTERIAL STRAINS (collection pictures-pdf)

Name

Section

Growth conditions

Acaryochloris marinaNBRC 102967 (MBIC11017)

I

Medium Acaryochloris (medio 963, NITE)

Anabaena azollaeAS-DS

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Anabaena cylindricaATCC 29414

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Anabaenasp. ATCC 33047

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Anabaenasp. PCC 6411 (ATCC 27898)

IV

BG110 +/- Nitrogen source (nitrate or ammonium). No fix N2 in oxic conditions.

Anabaena sp. PCC 7119 (ATCC 29151)

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Anabaenasp. PCC 7120 (ATCC 27893)

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Anabaenavariabilis ATCC 29413 (PCC 7937)

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Anabaenasp. Universidad de Leningrado 458 (PCC 7118)

IV

BG110 +/- Nitrogen source (nitrate or ammonium). No fix N2 in oxic conditions.

Anabaenasp. Universidad de Tokio M131

IV

BG110 +/- Nitrogen source (nitrate or ammonium). No fix N2 in oxic conditions.

Anabaena variabilisATCC 29413 - FD

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Anabaena variabilisATCC 29413 - P9

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Anacystis nidulansL-1402-1

I

BG11.

Arthrospira maxima

III

BG11C.

Calothrix (Tolypothrix)sp. PCC 7601

(Fremyella diplosiphon UTEX 481)

IV

BG110 +/- Nitrogen source (nitrate or ammonium). Fix N2 in anoxic conditions.

Calothrix sp. PCC 9327

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Chlorogloeopsissp. PCC 6912 (ATCC 29193)

V

BG110 +/- Nitrogen source (nitrate or ammonium).

Chroococcidiopsis thermalis PCC 7203

(Myxosarcina chroococcoides)

II

BG110 +/- Nitrogen source (nitrate or ammonium). Fix N2 in anoxic conditions.

Chroococcidiopsis sp. PCC 9317

II

BG11

Cyanothece sp. PCC 7425

I

BG110 +/- Nitrogen source (nitrate or ammonium). Fix N2 in anoxic conditions.

Cyanothece sp. PCC 9318

I

BG11

Dermocarpasp. PCC 7437 (Stanieria) (ATCC 29371)

II

BG11

Fischerella muscicola PCC 73103

V

BG110 +/- Nitrogen source (nitrate or ammonium).

Fischerella muscicola PCC 7414

V

BG110 +/- Nitrogen source (nitrate or ammonium).

Fischerella muscicolaUTEX 1829

V

BG110 +/- Nitrogen source (nitrate or ammonium).

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). Fix N2 in anoxic conditions.

Nodularia chucula

IV

BG110 +/- Nitrogen source (nitrate or ammonium)

Nostoc ellipsosporum(Univ. de Göttingen B1453-79)

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Nostoc paludosum  PCC 9206

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Nostoc sp. ATCC 29133

(PCC 73102) N. punctiforme

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Nostocsp. 78-IIA-E (ATCC 43237)

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Nostocsp. caquena

IV

BG110 +/- Nitrogen source (nitrate or ammonium)

Nostocsp. llaita

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Nostocsp. PCC 6705 (ATCC 29131)

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Nostocsp. PCC 6719 (ATCC 29105)

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Nostocsp. PCC 6720 (ATCC 27899)

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Nostocsp. PCC 7107 (ATCC 29150)

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Nostocsp. PCC 7413 (ATCC 29106)

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Nostoc sp. PCC 9201

IV

BG110 +/- Nitrogen source (nitrate or ammonium)

Nostoc sp. PCC 9202

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Nostocsp. PCC 9203

IV

BG110 +/- Nitrogen source (nitrate or ammonium)

Nostoc sp. PCC 9204

IV

BG110 +/- Nitrogen source (nitrate or ammonium)

Nostoc sp. PCC 9329

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Nostoc sp. PCC 9331

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Oscillatoria sp. PCC 9325

III

BG11

Phormidium laminosum (Agardh) Gom. OH-1-pC11

III

BG11. Growth at 45°C

Phormidium persicinum 80.79

III

Medium f/2

Pseudanabaenasp. PCC 6903 (ATCC 27190)

III

BG11

Pseudanabaena sp. PCC 9330

III

BG11

Scytonemasp. PCC 7110 (ATCC 29171)

IV

BG110 +/- Nitrogen source (nitrate or ammonium).

Spirulinasp. PCC 6313  (ATCC 29542)

III

BG11/ASNIII (1:1)

SynechococcusA6

I

BG11.

SynechococcusM4-

I

BG11.

Synechococcussp. PCC 6301 (ATCC 27144)  “S. leopoliensis

I

BG11

Synechococcussp. PCC 7202 (ATCC 29140)

I

BG11 + 10% ASN-III.

Synechococcussp. PCC 7335

I

ASN-III. Fix N2 in anoxic conditions.

Synechococcussp. PCC 7942  (Anacystis nidulans R2)

I

BG11

Synechococcus sp. PCC 9321

I

BG11

Synechocystissp. PCC 6308 (ATCC 27150)

I

BG11

Synechocystissp. PCC 6803 (ATCC 27184)

I

BG11

Synechocystis sp. PCC 9320

I

BG11

Synechocystis sp. PCC 9322

I

BG11

Synechocystis sp. PCC 9323

I

BG11

Thermosynechococcus elongatusBP-1

I

BG11. Growth at 45°C.


2. MICROALGAE

INTRODUCTION

Microalgae are eukaryotic organisms. They are (with numerous exceptions) aquatic organisms that (with frequent exceptions) are photosynthetic, oxygenic autotrophs that are typically smaller and less structurally complex than land plants. Algae are not a monophyletic group of organisms in which, depending on the different authors, different phyla or divisions can be distinguished: Glaucophyta, Euglenophyta, Cryptophyta, Haptophyta, Dinophyta, Ochrophyta, Rhodophyta and Chlorophyta. The CBCS microalgae collection contains members of different divisions.

GROWTH CONDITIONS

The culture media used in the CBCS for microalgae growth are, in many cases, specific for each microalga strain. In general, microalgae are maintained in solidified medium with bacto-agar 1%, or in liquid medium.
Microalgae are grown in phototrophy conditions at 25°C and low light intensity.
 

PHYLA OR DIVISION

 

Photosynthetic pigments

Storage products

Cell covering

Glaucophyta

Chlorofill a, phycocyanin, allophycocyanin, phycoerythrin, b-carotene, xanthophylls

Starch

Some cellulosic

Euglenophyta

Chlorofill a and b,

b-carotene, other carotenes and xanthophylls

Paramylon

Proteinaceous pellicle beneath plasma membrane

Cryptophyta

Chlorofill a and c phycocyanin, phycoerythrin, allophycocyanin, a, b-carotene, xanthophylls

Starch

Proteinaceous pellicle beneath plasma membrane

Haptophyta

Chlorofill a and c,

 b-carotene, xanthophylls

Chrysolaminaran

CaCO3 scales common

Dinophyta

Chlorofill a and c,

 b-carotene, xanthophylls

Starch

Cellulosic plates in vesicles beneath plasma membrane

Ochrophyta (diatoms, chrysophyta, brown algae…)

 

 

(eustigmatophytes)

Chlorofill a and c,

 b-carotene, xanthophylls 

 

 

Chlorofill a,

 b-carotene, xanthophylls 

Chrysolaminaran, lipids

 

 

 Chrysolaminaran

Some naked, some with silica/organic scales; cellulose, alginates in some

 

Present, but unknown composition

Rodophyta

Chlorofill a, phycocyanin, allophycocyanin, phycoerythrin, a, b-carotene, xanthophylls

Floridean starch

Cellulose, sulfated polysaccharides; some calcified

Chlorophyta

Chlorofill a and b,

b-carotene, other carotenes and xanthophylls

Starch

Wall of cellulose/other polymers; scales on some; some naked; some calcified

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

LIST OF MICROALGAE STRAINS (collection pictures-pdf)

Name

Division

Growth conditions

Astasia longa1204-17a

Euglenophyta

Euglena Medium

Chlamydomonas augustae(C. nivalis o Chloromonas reticulata) CCAP11/51,

SAG 26.86

Chlorophyta

SUEOKA. Nit+

Chlamydomonas euryale25-89

Chlorophyta

SUEOKA.  Nit -

Chlamydomonas reinhardtii 21gr (+)

Chlorophyta

SUEOKA. Nit+

Chlamydomonas reinhardtii  137c (+)

Chlorophyta

SUEOKA. Nit -

Chlamydomonas reinhardtii  CC124 (-)

Chlorophyta

SUEOKA. Nit -

Chlamydomonas reinhardtii  CC621 (-)

Chlorophyta

SUEOKA. Nit -

Chlamydomonas reinhardtii CF10 (-)

Chlorophyta

SUEOKA. Nit+

Chlamydomonas reinhardtii  CF11 (-)

Chlorophyta

SUEOKA. Nit -

Chlamydomonas reinhardtii  CF14 (+)

Chlorophyta

SUEOKA. Nit+

Chlamydomonas reinhardtii CW15-325 (+)

Chlorophyta

SUEOKA. Nit + Arg -

Chlamydomonas reinhardtii CW15 (-)

Chlorophyta

SUEOKA. Nit +

Chlamydomonas reinhardtii 704 (+)

Chlorophyta

SUEOKA. Nit +

Chlorella sp. CCAP 211/84

Chlorophyta

EG:JM. Nit +

Chlorella fusca(C. emersonii) 211-8b

Chlorophyta

SUEOKA. Nit +

Chlorella salina8-86

Chlorophyta

f/2 medium

Chlorella sorokiniana211-32

Chlorophyta

SUEOKA. Nit +

Chlorella stigmatofora 9.86

Chlorophyta

 f/2 medium

Chlorella vulgaris 9-88 (UAM 101)

Chlorophyta

SUEOKA. Nit +/-

Chlorella zofingiensis211/14

Chlorophyta

SUEOKA. Nit +/-

Chlorococcum citriforme62.80

Chlorophyta

SUEOKA. Nit +

Chromera velia CCAP 1602/1

Chromerida

 f/2 medium

Coelastrum proboscideum

(var. dilatatum) 217-2

Chlorophyta

SUEOKA. Nit +

 

Coelastrum proboscideum

(var. gracile) 217-3

Chlorophyta

SUEOKA. Nit +

 

Cryptomonas sp.06/0101

Cryptophyta

 f/2 medium

Cyanidium caldarium16-91

Rhodophyta

Cyanidium medium

Cyanophora paradoxa

Glaucophyta

BG11 (2 mM NaNO3)

Dunaliella salina

Chlorophyta

Dunaliella medium

Euglena gracilisSAG 5-9

Euglenophyta

Euglena medium

Euglena gracilis SAG 5-15

Euglenophyta

Euglena medium

Euglena gracilis SAG 5-25

Euglenophyta

Euglena medium

Galdieria sulphuraria 74W

Rhodophyta

Cyanidium medium

Haematococcus lacustris192.80

Chlorophyta

H medium

Haematococcus lacustrisATCC 30453

Chlorophyta

H medium

Haematococcus pluviales CCAP 34/8

Chlorophyta

H medium

Haematococcus pluvialisSAG 192.80

Chlorophyta

H medium

Haematococcus pluviales SAG 34-1b

Chlorophyta

H medium

Heterocapsa sp. 09/0601

Dinophyta

 f/2 medium

Isochrysis galbana 05/0401

Ochrophyta

 f/2 medium

Micromonas pusilla RCC 827

Chlorophyta

Acaryochloris medium

Monoraphidium braunii202.7d

Chlorophyta

SUEOKA. Nit +

Monoraphidium circinale

Chlorophyta

 f/2 medium (0,25x)

Muriellopsissp. IBVF-L4

Chlorophyta

SUEOKA. Nit +

Nannochloropsis gaditana

Ochrophyta

 f/2 medium

Nannochloris atomus SAG 14.87

Chlorophyta

 f/2 medium

Nannochloris maculata 251/3

Chlorophyta

 f/2 medium

Nannochloropsis oculata 38.85

Ochrophyta

 f/2 medium

Nannochloropsis salina40.85

Ochrophyta

 f/2 medium

Navicula pelliculosa1050-3

Ochrophyta

Especific

Neochloris oleoabundans UTEX 1185

Chlorophyta

BG11

Neospongiococcum (Chlorococcum) gelatinosum 64.80

Chlorophyta

SUEOKA. Nit +

Nephroselmis olivacea CCAP 1960/4B

Chlorophyta

3N-BBM+V (CCAP)

Ochromonas danica933-7

Ochrophyta

Euglena medium

Ostreococcus tauri RCC745

Chlorophyta

f/2 medium

Phaeodactylum tricornutum 07/0502

Ochrophyta

 f/2 medium

Phaeodactylum tricornutum CCAP 1055/1

Ochrophyta

 f/2 medium

Porphyridium purpureum1380-1a

Rhodophyta

 f/2 medium

Poterioochromonas malhamensis CCAP 933/1C

Ochrophyta

Euglena medium

Pycnococcussp. RCC245       

Chlorophyta

Acaryochloris medium

Pycnococcus provasoliiCCAP 190/1

Chlorophyta

 f/2 medium

Rhodomonas baltica 06/0301

Cryptophyta

 f/2 medium

Rhodomonas salina

Cryptophyta

 f/2 medium

Scenedesmus vacuolatus  SAG 211-15

Chlorophyta

SUEOKA. Nit +

Spirogyra sp SAG 170.80

Streptophyta

BG11

Tetracystissp.

Chlorophyta

SUEOKA. Nit +

Tetraselmis chuii 8-6

Chlorophyta

 f/2 medium.

Tetraselmis chuiRCC128      

Chlorophyta

Acaryochloris medium

Tetraselmis suecica03/0203

Chlorophyta

 f/2 medium.


3 Escherichia coli

INTRODUCTION

Escherichia coli is an enterobacteria very used in molecular biology thanks to its high growth rate, the knowledge of its genome sequence and the posibility to introduce external DNA into the cells by transformation, transduction or electroporation. The CBCS E. coli collection is used by IBVF members in DNA recombinant works.

List of E. coli strains

CEPA

Genotipo

E. coliBL21 (DE3)

F- ompT (lon) hsdS(rB-mB-, una E. coli B) con DE3.

E. coliBL21 (DE3) lacIq

F´::Tn10 proA+B+ lacIq D(lacZ)M15/ ompT (lon) hsdS (rB-mB-, una E. coli B) con DE3.

E. coli BL21 (DE3) pLysS

F- ompT hsdSB (rB-mB-) gal dcm (DE3) pLysS (CmR)

E. coliBL21 (DE3)(pIZ227)

pIZ227 porta, junto con lacIq, un gen de una lisozima específica de la polimerasa T7. Cmr.

E. coliBL21 CodonPlus(DE3)-RIL

E. coliB F- ompT (lon) hsdS(rB-mB-) dcm+ Tetr gallHte (argU ileY LeuW Cmr)     -argU, AGA/AGG; IleY, AUA; LeuW, CUA-

E. coli BL21 Rosetta(DE3)pLys

F-ompThsdSB(rB-mB-) gal dcm (DE3)pLysRARE (CmR) (tRNAs para AGG, AGA, AUA, CUA, CCC y GGA en pLys)

E. coli BL21-AI

F- ompT hsdSB (rB-mB-)gal dcm araB::T7RNAP-tetA  lon (tetR)

E. coli BL21-T1R

F- ompT hsdSB(rB-mB-) gal dcm tonA  

E. coli C41(DE3)

F ompT hsdSB (rB- mB-)gal dcm (DE3)

E. coli C43(DE3)

F ompT hsdSB (rB- mB-)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 (rK-rK+) recA1 girA96 (Nalr) 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(rB-mB-) leu supE44 ara14 galK2 lacY1 proA2 rpsL20 xyl-5 mtl-1 recA13 mcrB

E. coliJM109

traD36 lacIq D(lacZ)M15 proA+B+/e14-(McrA-) D(lac-proAB) thi gyrA96(Nalr) endA1 hsdR17 (rK-mK+) 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 (CmR)

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(KmR, lacIq)

E. coliXL1-Blue

 

F´::Tn10 proA+B+ lacIq D(lacZ)M15/recA1 endA1 gyrA96(Nalr) thi hsdR17 (rK-mK+) supE44 relA1 lac

E.coli Solu (BL21)

F- ompT hsdSB (rB-mB-) 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(Strr) hsdR2 (rK- mK+) mcrA mcrB1

 

4. CULTURE MEDIA

 

BG11MEDIUN (Rippka et al, 1979)

Na2CO3       

0.2 mM   

 H3BO3

46 µM           

MgSO4.7H2O            

0.3 mM

MnCl2.4H2O            

9.1 µM

CaCl2.2H2O            

0.24 mM

Na2MoO4

1.6 µM

K2HPO4

0.2 mM

ZnSO4.7H2O            

0.8 µM

ácido cítrico  

28.5 µM

CuSO4.5H2O            

0.3 µM

Ferric-ammonium citrate (17% Fe)

6 mgl-1

CoCl2.6H2O            

0.2 µM

Na2-EDTA

2.4 µM

 

 

Medium is prepared from a 100x-fold stock without K2HPO4 and nitrogen source, which are added prior autoclaving. For solid medium, Bacto-agar is used at 1% w/v. Agar is autoclaved in half volume, separated from nutrients. 

 

BG11 MEDIUM

Medium BG110 supplemented with NaNO3 at 17.6 mM.

 

BG11C MEDIUM

Medium BG110 supplemented with NaNO3 at 17.6 mM and NaHCO3 at 10 mM.

 

ASN-III MEDIUM (Rippka et al, 1979)

NaCl                          25 g/l

Ferric-ammonium citrate (17% Fe)       0.003 g/l

MgSO4.7H2O            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

Trace metals (A5+Co)                      1 ml/l

K2HPO4                   0.02 g/l

 

NaNO3                            0.75 g/l

 

CaCl2.2H2O              0.5 g/l

 

Citric acid              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

 

SUEOKA MEDIUM

 

KH2PO4                      8.3 mM

K2HPO4                      5.3 mM

MgSO4.7H2O              0.25 mM

CaCl2.2H2O                0.133 mM

Traces HUTNER            1x

NH4Cl                         9.35 mM

For solid medium, Bacto-agar is used at 1% w/v. Agar is autoclaved in half volume, separated from nutrients. 

Traces Hutner 200x:

  1. Dissolve 12.7 g de EDTA-Na.2H2O in 250 ml of water.
  2. Dissolve 1 to 1 in this order, heating previously at 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

  1. Mix both solutions.
  2. Heat up to 100ºC and allow cooling up to 80-90ºC.
  3. Adjust pH at 6.8 with KOH 20% (Tª always >70ºC).
  4. Add water up to 1 liter and keep in dark for 2 days.

 

H MEDIUM

Sueoka medium supplemented with 1.5 g/l proteose peptone.

 

CYANIDIUM MEDIUM

 

K2HPO4

0.3 g

CaCl2.2H2O            

0.02 g

MgSO4.7H2O

0.3 g

KNO3

3.0 g

FeSO4.7H2O

0.003 g

H2SO4

1.8 g

Trace metals (A5+Co)

1 ml/l

Water (up to 1 liter)

 

 

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

 

EUGLENA MEDIUM

 

Sodium acetate                   1 g

Beef extract                         1 g

Bacto-triptone                    2 g

Yeast extract                      2 g

CaCl2.2H2O                   10 mg

Water (up to 1 liter)

For solid medium, Bacto-agar is used at 1% w/v. 

 

Guillard's f/2 MEDIUM

Add to 998 ml of filtered natural sea water 1 ml of stocks 1 and 3. Autoclave and add, once cooled, 200 µl of stock 2 (sterilized by filtration 0,2 µm).

Stock 1. Trace metals

 

Stock 2. Vitamins.

 

Stock 3.

EDTA-Na2               4.16 g

Nicotinic acid                    1 mg/ml

KNO3             15 g

FeCl36H2O              3.15 g

Para-aminobenzoic acid   1 mg/ml

NaH2PO4     1.13 g

CuSO45H2O            0.01 g

NaHCO3                      0.5 mg/ml

Water

(up to 100 ml)

ZnSO47H2O          0.022 g

Thiamine HCl                0.5 mg/ml

 

CoCl26H2O             0.01 g

Biotin                          0.1 mg/ml

 

MnCl24H2O            0.18 g

Vit. B12                    0.035 mg/ml

 

Na2MoO42H2O      0.006g

 

 

Water (up to 1 liter)

 

 

 

DUNALIELLA MEDIUM

 

Base solution (4x)

 

Microelements (1000x)

KCl                                      20 mM

MnCl2.4H2O                             7 mM

MgCl2.6H2O                         20 mM

CuCl2.2H2O                              1 mM

Na2SO4                                20 mM

ZnCl2                                        1 mM

CaCl2.2H2O                         0.8 mM

CoCl2.6H2O                              1 mM

 

Fe3+-EDTA (1000x)

 FeCl3.6H2O                2 mM

 NaEDTA                    5 mM

 

Others (final concentrations)

NaNO3                       5 mM

NaCl                           0.5 M (solid medium)/3 M (liquid medium)

KH2PO4                     0.2 mM

NaHCO3                    50 mM

 

Sterilize by autoclaving (KH2PO4 and NaHCO3 separated). For solid medium, Bacto-agar is used at 1% w/v. Agar is autoclaved in half volume, separated from nutrients.

 

NAVICULA MEDIUN

 

NaNO3                             1.18 mM

K2HPO4                          77 mM

Na2SiO3                          0.86 mM

Triptona                           1 g/l

BG11, 100x-fold stock          2x

For solid medium, Bacto-agar is used at 1,5% w/v. Agar is autoclaved in half volume, separated from nutrients.  

 

ACARYOCHLORIS MEDIUM

 

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

Thiamine-HCl      

0.2 mg

Biotin

0.0015 mg

Vitamin B12

0.0015 mg

Natural sea water         

Up to 1000 ml