Vitamin Mixtures – Plant Tissue Culture Protocol

• Preparation of Solutions from Packaged Powder
  
• Media Preparation using Vitamin Solutions
  
• Vitamin Mixtures Composition Table

 

Preparation of Solutions from Packaged Powder

Powdered vitamin mixtures are hygroscopic and must be protected from atmospheric moisture. The entire contents of each package should be used immediately after opening. The basic steps for preparing 1000X concentrated solutions with our vitamin mixtures are listed below:

1. Measure out 70% of the final required volume of deionized-distilled water (e.g. 70 mL for a final volume of 100 mL).
   
2. While stirring the water add the powdered vitamin mixture. Stir until completely dissolved. Increasing the pH and/or warming solution (35-37 °C) may be required.
3. Rinse the original container with a small volume of water to remove traces of the powder. Add to the solution in Step No. 2.
4. Add additional water to bring the medium to the final volume.
5. The resulting 1000X concentrated solution should be used at a concentration of 1 mL/L of medium.
6. Follow the same steps to prepare a 100X concentrated solution and use at 10 mL/L of medium.

Media Preparation Using Vitamin Solutions

Prepared Vitamin Solutions:

1. The vitamin solutions are sterile filtered through a double 0.2 µm filtration unit and are ready for use.
2. Vitamin solutions should be added at a concentration of 1 mL/L of medium to prepare the final recommended concentration of vitamins in the medium. (Except K3129, Kao and Michayluk vitamin solution, which should be added at a concentration of 10 mL/L of medium.)

Materials Not Provided

• BASAL MEDIA AND ADDITIVES AS REQUIRED
• Deionized tissue culture grade water (W3500)
• 1 N Hydrochloric Acid (H9892)
• 1 N Sodium Hydroxide (S2770)

VITAMIN POWDERS AND SOLUTIONS ARE FOR LABORATORY USE ONLY. NOT FOR DRUG, HOUSEHOLD OR OTHER USES.

Storage

Store powdered vitamin mixtures and solutions in a refrigerator at 0-5 °C (except  K3129, Kao and Michayluk vitamin solution, which should be stored at -0 °C). Precipitation in vitamin solutions may occur during storage. This can be redissolved by warming the solution in a water bath (35-37 °C) for a short period of time.

Vitamin Mixtures Composition Table

G1019 – Gamborg's vitamin solution (1000X)
K3129 – Kao and Michayluk vitamin solution (100X)
M7150 – Murashige and Skoog vitamin powder
M3900 – Murashige and Skoog vitamin solution (1000X)
S3766 – Schenk and Hildebrandt vitamin powder

Component (mg/L)*	G1019	K3129	M7150	M3900	S3766
p-Aminobenzoic acid		0.02
L-Ascorbic acid		2.0
Biotin		0.01
D-Calcium pantothenate		1.0
Choline chloride		1.0
Choline-H2 citrate
Folic acid		0.4
Folic acid, Ca
Glycine			2.0	2.0
myo-Inositol	100.0	100.0	100.0	100.0	1000.0
Nicotinamide (Niacinamide)		1.0
Nicotinic acid	1.0		0.5	0.5	5.0
Orotic acid
Putrescine • 2HCl
Pyridoxine • HCl	1.0	1.0	0.5	0.5	0.5
Pyridoxamine • 2HCl
Riboflavin		0.2
Thiamine • HCl	10.0	1.0	0.1	0.1	5.0
Thymine
Vitamin A (Retinol)		0.01
Vitamin B12		0.02
Vitamin D (Cholecalciferol)		0.01
*Concentrations listed are final concentrations when used at recommended dilution