Medium Recipes
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Cryoprotectants Mint Potato

 

Stock Solutions
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Macroelement
Micronutrients
Iron 37.3 mg/L of NaEDTA and 27.8 mg/L of FeSO4 (7H2O) H3BO 0.62 g/L
NH4NO3 1.65 g/L CoCl2 2.5 mg/L
MgSO4 370 mg/L CuSO4 2.5 mg/L
CaCl2 440 mg/L ZnSO4 0.86.g/L
KPH2O4 170 mg/L NaMo04 25 mg/L
KNO3 1.9 g/L MnSO4 1.69 g/L
    KI 83 g/L

 

Vitamins
MS Vitamins
MS-G 0.08 g/L Thiamine and
20 g/L Myoinositol		 Glycine 0.2 g/L
Myoinositol 10 mg/L Myoinositol 10 g/L
Folic Acid 20 mg/L Nicotinic Acid 50 mg/L
    Pyridoxine HCl 50 mg/L
    Thiamine HCl 10 mg/L


Supplemental Vitamins for Potato Liquid Medium
Growth Regulators
Biotin 25 mg/250 ml Kinetin  
Thiamine 25 mg/250 ml IAA 12.5 mg/250 ml
Pyridoxine HCL 25 mg/250 ml GA3 12.5 mg/250 ml
Nicotinic Acid 50 mg/250 ml Zeatin 3.125 g/62.5 ml
Riboflavin 12.5 mg/250 ml IBA 10.16 mg/50 ml
Glycine 75 mg/250 ml BA 5 mg/100 ml
Pantothenic Acid 250 mg/250 ml    
Ascorbic Acid 50 mg/250 ml    
Cyanocobalamin (B12) 1.5 mg/250 ml    
Pour 10 ml per vessel, Freeze
    

 

Mint Medium
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Total amount = 500 ml

Mint Growth Media
Mint Recovery Media
Ingredients from Stock Solutions
 KNO3 10 ml
KNO3 10 ml NHNO3 10 ml
NHNO3 10 ml MgSO4 5 ml
MgSO4 5 ml CaCl2 5 ml
Cacl2 5 ml KH2PO4 5 ml
KH2PO4 5 ml Iron 5 ml
Iron 5 ml Micronutrients 5 ml
Micronutrients 5 ml MS Vitamins 5 ml
MS vitamins 5 ml Sucrose 15 g
Sucrose (3% use table sugar) 15 g Benzyladenine (BA) 5 ml
pH 5.7*   Indole 3 Butyric Acid (IBA) 0.25 ml
Agar (.3%) 1.5 g pH 5.7*
Gelrite (.125%) .625 g Agar 1.5 g
    Gelrite (.125%) 0.625
*pH prior to addition of gelling agent

Reference:

Murashige, T. and F. Skoog. 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiology Plantarum 15:473-497

 

Potato Medium
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Total amount = 500 ml

Solid Medium
Liquid Medium
Ingredients from Stock Solutions
Ingredients from Stock Solutions
KNO3 10 ml KNO3 5 ml
NHNO3 10 ml NHNO3 5 ml
MgSO4 5 ml MgSO4 5 ml
CaCl2 5 ml KH2PO4 5 ml
KH2PO4 5 ml CaNO3 .15 g
Iron 5 ml Iron 5 ml
Micronutrients 5 ml Micronutrients 5 ml
Sucrose (2% use table sugar 10 g Myoinositol 5 ml
pH 5.7* Folic acid .5 ml
Agar (.8%) 4 g Supplemental Vitamins 5 ml
MS-G 5 ml
Kinetin 2.5 ml
IAA .01 ml
Sucrose (3% use table sugar) 15 g
pH 5.65  
125 ml placed in container (13 cm tall x 11.5 cm round)  
*pH prior to addition of gelling agent
See References Below

 

MSTo (500 ml)
sweet potato plant and flowers
KNO3 10 ml
NHNO3 10 ml
MgSO4 5 ml
CaCl2 5 ml
KH2PO4 5 ml
Iron 5 ml
Micronutrients 5 ml
mixed potato tubers
MS Vitamins 5 ml
Disaccharides 15 g
IAA 5 ml
GA3 2 ml
Zeatin 5 ml
pH 5.7  

Reference:

Murashige, T. and F. Skoog. 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiology Plantarum 15:473-497

Schafer-Menuhr, A. 1996. Refinement of cryopreservation techniques for potato. DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen Gmbh. Braunschweig, Germany.

Towill, L.E. 1983. Improved survival after cryogenic exposure of shoot-tips derived from in vitro plantlet cultures of potato. Cryobiology 20: 567-573.


Cryoprotectant and Retrieval Solutions
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2% Sucrose

MS medium is made with modification based on which plant is being preserved. Sigma Grade II sucrose is added to final concentration of 2%.

0.3M Sucrose

MS medium is made with modification based on which plant is being preserved. Sigma Grade II sucrose is added to final concentration of .3M.

0.4M Sucrose + 2M Glycerol

MS medium is made with modification based on which plant is being preserved. Sigma Grade II sucrose and glycerol are added to final concentration of .4M and 2M, respectively.

1.2M Sucrose

MS medium is made with modification based on which plant is being preserved. Sigma Grade II sucrose is added to final concentration of 1.2M.

PVS2 (50 ml)

media preparation tools
KNO3 1 ml
NHNO3 1 ml
MgSO4 0.5 ml
CaCl2 0.5 ml
KH2PO4 0.5 ml
Iron 0.5 ml
Micronutrients 0.5 ml
MS Vitamins 0.5 ml
DD H20 19.5 ml
pH 5.7*
Add to Solution
Glycerol 11.9 ml
Ethylene 6.8 ml
Autoclave
Add to Solution under sterile hood
DMSO by filter sterilization 6.8 ml

Reference:

Sakai, A., S. Kobayashi and I. Oiyama. 1991. Survival by vitrification of nucellar cells of navel orange (Citrus sinensis var. brasiliensis Tanaka) cooled to -196°C.
J. Plant Physiol. 137:465-470.

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