Kamaishu Bio recommended today to read - MS medium mother liquid configuration and preservation

Instruments and appliances

1. Instruments: all kinds of balances, magnetic stirrers, refrigerators;

2, utensils: beakers, volumetric flasks, measuring cylinders, reagent bottles, labels;

3. Reagents: 95% alcohol, 0.1-1N NaOH, 0.1-1NHCl, various inorganic substances, organic substances and distilled water required for preparation of MS medium.

Method step

(1) Preparation of mother liquor

The mother liquor is a concentrate to be prepared for the medium, and is generally formulated to be a solution 10 to 100 times higher than the desired concentration.

Advantages: (1) to ensure the accuracy of the composition of each substance.

(2) Quickly move when it is easy to configure.

(3) It is convenient for cryopreservation.

1, MS mass element mother liquor (10X)

A 10 liter amount was dissolved in 1 liter of distilled water. One liter of medium was taken to take 100 ml of the mother liquor.

chemical

1 liter

10 liters

1

NH4NO3

1650 mg/L

16.5g

2

KNO3

1900 mg/L

19.0g

3

CaCl2·2H2O

440 mg/L

4.4g

4

MgSO4·7H2O

370 mg/L

3.7g

5

KH2PO4

170 mg/L

1.7g

2. MS trace element mother liquor (100X)

A 10 liter amount was dissolved in 100 ml of distilled water. One liter of medium was taken to take 10 ml of the mother liquor.

chemical

1 liter

10 liters

1

MnSO4·4H2O

(MnSO4·H2O)

22.3 mg/L

(21.4 mg/L)

223mg

2

ZnSO4·7H2O

8.6 mg/L

86mg

3

CoCl2·6H2O

0.025mg/L

0.25mg

4

CuSO4·5H2O

0.025 mg/L

0.25mg

5

Na2MoO4·2H2O

0.25 mg/L

2.5mg

6

KI

0.83 mg/L

8.3 mg

7

H3BO3

6.2 mg/L

62 mg

Note: CoCl2·6H2O and CuSO4·5H2O can be weighed in 10 times (0.25 mg×10=2.5 ​​mg) or 100 times (25 mg), and then fixed to 100 ml of water, each time taking 10 ml or 1 ml (including An amount of 0.25 mg) was added to the mother liquor.

3, MS iron salt mother liquor (100X)

A 10 liter amount was dissolved in 100 ml of distilled water. One liter of medium was taken to take 10 ml of the mother liquor.

chemical

1 liter

10 liters

Na2·EDTA

37.3 mg/L

373 mg

FeSO4·7H2O

27.8 mg/L

278 mg

Note that when preparing, the two components should be dissolved in a small amount of distilled water, wherein the EDTA salt is difficult to completely dissolve, and can be heated appropriately. When mixing, first take a volumetric flask (beaker), then shake the other component one by one to produce a dark yellow solution, and finally set it to a brown reagent bottle.

4, MS organic mother liquor (100X)

A 10 liter amount was dissolved in 100 ml of distilled water. One liter of medium was taken to take 10 ml of the mother liquor.

chemical

1 liter

10 liters

1

niacin

0.5 mg/L

5mg

2

Pyridoxine hydrochloride (VB6)

0.5 mg/L

5mg

3

Thiamine hydrochloride (VB1)

4

Inositol

100 mg/L

1 g

5

Glycine

2 mg/L

20mg

5, growth regulator

Prepared separately, at a concentration of 1-5 mg/ml (0.5-1 mg/ml in the book), usually formulated at 4 mg/ml.

Precautions when preparing the mother liquor:

1 Some ions are prone to precipitation, which can be first dissolved in a small amount of distilled water, and sequentially mixed in the order of formulation;

2 Distilled water or double distilled water must be used to prepare the mother liquor;

3 pharmaceutical applications are chemically pure or analytically pure;

4 When auxin is dissolved, it can be dissolved with a small amount of 0.5 –1N NaOH or 95% alcohol, and the dissolved mitogens are dissolved by heating with 0.5–1N HCl.

(2) Preservation of mother liquor

1. Bottling: The prepared mother liquor is separately filled into the reagent bottle, and the label is attached, indicating the name, concentration factor and date of each mother liquor. Note that the easily decomposable, oxidized solution is stored in a brown bottle.

2, storage: 4 ° C refrigerator.

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