MS media (Murashige – Skoog) composition and preparation

Table Of Contents


The composition of the growing medium is crucial in every experiment containing living organisms. The need for a well-balanced plant medium arose at the beginning of hydroponic cultivation and plant tissue cultures. The first recipes for plant tissue cultures are connected with great names of plant in vitro – Hildebrandt and others (e.g. Hildebrandt et al., 1946). Important research in the field of plant media was done in the 60s and 70s, probably the most important publication is Murashige and Skoog (1962). This medium was originally designed for nicotiana (Tabaco) calli but it is used in altered forms for other plants – for instance, omnipresent Arabidopsis thaliana.

Naturally, different plants require different compositions of media but MS-based media are widely used for many species nowadays. However, there are also many exceptions as plants are diverse organisms – for example, Knudson media for orchids (Knudson, 1951; Sigma-Aldrich-a). The difference in composition with common alternatives is mentioned below. To sum it up, they differ in quantities of macronutrients (for instance, some plants demand more nitrogen or phosphorus) but also in presence of micronutrients – that determinates some media only for short-term cultivation.


  • The principal component of In-vitro plant cultivation.
  • Plant tissue culture media like calli induction, shoot regeneration, and root formation.
  • For the germination of seeds of different plant species like Arabidopsis thalianaNicotiana benthamiana, under sterile growth conditions.


The compounds used in the media can be divided into the following categories:

  • Basal salts (macro and micronutrients)
  • Vitamins (and organic essentials and beneficial)
  • Buffering substances, carbon resource (sucrose) and
  • Reinforcing agent (none in liquid, agar, or phytagel in solid)

For making MS, you can buy different products – a full-fledged MS (Thermofisher), only basal salts (both or separate micro and macronutrients), basal salts with vitamins, and many other combinations (Sigma-Aldrich-b).

Here we are describing the total composition, so it can be made from basic reagents or complemented to some commercial products. Sometimes the composition of the medium is cross-over – for example, Murashige-Skoog medium with Gamborg vitamins.

The tables below with a comparison of MS to other used media for In-vitro plant/tissue cultivation (adapted from Misawa 1994).

Basal salts

Components (mg/l)Murashige - Skoog (1962)White (1963)Gamborg (1968)Heller (1953)Schenk - Hildebrandt (1972)Nitsch - Nitsch (1967)Kohlenbach - Schmidt (1975)
KNO31 900803 000-2 500125950
MnSO4.4H2O22.3710 (1 H2O)0.1102525

*For Arabidopsis thaliana, ammonium nitrate is sometimes omitted.

Table 1. Comparision of various plant culture media composition.

Vitamins (essentials and beneficial)

Components (mg/l)Murashige - Skoog (1962)White (1963)Gamborg (1968)Heller (1953)Schenk - Hildebrandt(1972)Nitsch - Nitsch (1967)Kohlenbach - Schmidt (1975)
Myo-inositol100-100-1 000100100
Nicotinic acid0.50.51-0.555
Pyridoxine HCl0.50.11-
Thiamine HCl0.1-10.110150.50.5
Cysteine HCl-1-----
Folic acid-----0.50.5

*For Arabidopsis thaliana, ammonium nitrate is sometimes omitted.

Table 2. Vitamins used in various plant culture media.

Carbon source

Components (g/l)Murashige - Skoog (1962)White (1963)Gamborg (1968)Heller (1953)Schenk - Hildebrandt(1972)Nitsch - Nitsch (1967)Kohlenbach - Schmidt (1975)

*For Arabidopsis thaliana, ammonium nitrate is sometimes omitted.

Table 3. Carbon source used in  plant culture media.

Buffering substances

Tris, Tricine, MES, HEPES, PB-74, and CAPS have been used in tissue cultures for buffering. They differ in toxicity for particular plant species. MES is widely used because it has a good buffering capacity in mild acidic pH, exhibits low binding to nutrients, and exhibits low toxicity in most of the plants (except Brassica napus protoplasts – PB-74 is used there). (George et al., 2007).

0.5 g/l of MES is used.

Summary of MS composition

Category of componentsComponentsFor 1 l of media (mg)Total mass of components (mg)
MacronutrientsMgSO4.7H2O3704530 (2880**)
MacronutrientsCaCl2.2H2O 4404530 (2880**)
MacronutrientsKNO319004530 (2880**)
MacronutrientsNH4NO31650**4530 (2880**)
MacronutrientsKH2PO41704530 (2880**)
MicronutrientsNa2EDTA*37.3103.1 - 104
MicronutrientsMnSO4.4H2O22.3103.1 - 104
MicronutrientsZnSO4.7H2O8.6103.1 - 104
MicronutrientsCuSO4.5H2O0.025103.1 - 104
MicronutrientsCoCl2.6H2O0.025103.1 - 104
MicronutrientsKI0.83103.1 - 104
MicronutrientsH3BO36.2103.1 - 104
MicronutrientsNa2MoO4.2H2O0.25103.1 - 104
VitaminsMyo-inositol100103.1 - 104
VitaminsNicotinic acid0.5103.1 - 104
VitaminsPyridoxine HCl0.5103.1 - 104
VitaminsThiamine HCl0.1-1103.1 - 104
VitaminsGlycine2103.1 - 104
Carbon resourceSucrose3000030000
Buffering substanceMES500500
Reinforcing agent (one of those)Agar5000-100005000-10000
Reinforcing agent (one of those)Phytagel1500- 25001500- 2500

*EDTA chelates metal ions and prevents them from hydrolysis or immobilization.

**Ammonium nitrate can be omitted for Arabidopsis thaliana.

Table 4. Composition of MS medium.


Acidic solution (pH 5.7-5.8) is used in the original study (Murashige and Skoog, 1962).

Most known plant tissues require pH 5.2-5.8 (Skirvin et al., 1986).

The pH will be under constant change during the cultivation, so stable buffering conditions are essential for long-term cultivation. MES is commonly used because it buffers well between pH 5-7 (approximately) and it is not toxic to most plants species.

Medium is usually adjusted with NaOH/HCl. KOH can also be used instead of NaOH.


Basal salts


Buffering substance (MES)

Carbon resource (sucrose)

Reinforcing agent (agar)

NaOH/HCl for pH adjustment

Instruments and other requirements

  • Glass beaker
  • Weighing balance
  • Magnetic stirrer and pellet
  • pH meter
  • Measuring cylinders
  • Pipette
  • Bottles for autoclaving


  1. Using a weighing balance, prepare exact amounts of basal salts, vitamins, sucrose, MES, and agar (for solid medium).
  2. Note: If you prepare the MS from basic reagents, you can prepare stock powder of basal salts and vitamins (well-mixed). Then just weigh the total mass mentioned in the summary table.
  3. Prepare ¾ of the final volume of distilled water in a glass beaker.
  4. Put salts, vitamins, sucrose, and MES to dissolve (Do not add agar yet!). A magnetic stirrer can be used.
  5. After everything is dissolved, measure and adjust pH.
  6. Adjust the final volume using a measuring cylinder.
  7. Transfer the liquid to bottles for autoclaving and add agar (if you have more bottles then add a proportional amount of agar to every single bottle).
  8. Autoclave the bottles with media. Too long autoclaving causes degradation of some compounds.
  9. After autoclaving, let the bottles cool down a bit before pouring them into plates. Working with nearly boiling liquid is not pleasant. Moreover, the agar congeals in temperature 30-35 °C, so you can wait a bit after the autoclaving.
  10. Try to avoid working in a non-sterile environment – use a flow box.
  11. If you want to add hormones/antibiotics, add them to the cooling medium. (Five seconds rule is useful here. If you can hold your hand for five seconds or more on the bottle, you can add labile compounds.)
  12. Now you can pour the medium into plates. Mix the bottle before every pouring.

Preparation of solid media

For solid medium, use either agar or phytagel.

Agar: 5-10 g/l

Phytagel: 1.5-2.5 g/l


The universal storing condition is 4 °C. Store in this temperature the basal salts and vitamins stock, prepared liquid media, plates with solid MS.

Points to be noted

MS medium is a safe medium. The major problems could be connected with too long autoclaving (even caramelized sugar) and non-sterile conditions of work (sugar is an optimal nutrient for bacteria and fungi).

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