1/2 MS low Iron

1/2 MS low Iron kdorfman Tue, 02/15/2022 - 15:00

1/2 Murashige & Skoog, 1µM iron medium

MS 10x micronutrients is 100 µM FeSO4, so MS complete is 10 µM, 1/2 MS is 5µM

FeSO4 stock is 10 mM, which is 10,000x 1 µM

LPGM

LPGM kdorfman Wed, 08/31/2016 - 20:24

Lilly pollen growth medium

205 mM (7%) sucrose (See Sugars.)

1.6 mM H3BO3

0.1 mM CaCl2

15 mM MES

Either make fresh each day, or filter sterilize for all the labs.

Stock solutions:

150 mM MES

10 mM CaCl2

160 mM Borate

MS high salt

MS high salt kdorfman Mon, 02/03/2014 - 15:33

150 too high - try 100mM for F 2015

150 mM NaCl

10 g agar/L

Make regular MS, then add 0.03 mL 5M NaCl per mL medium

100 mM NaCl

0.02 mL 5M NaCl per mL medium

MS iron-free

MS iron-free kdorfman Thu, 05/23/2013 - 17:46

Murashige & Skoog 1µM iron medium

MS 10x micronutrients is 100 µM FeSO4, so MS complete is 10 µM

FeSO4 stock is 10 mM, which is 10,000x 1 µM

10 g agar/L

MS low iron

MS low iron kdorfman Thu, 05/23/2013 - 17:40

Murashige & Skoog 1µM iron medium

MS 10x micronutrients is 100 µM FeSO4, so MS complete is 10 µM

FeSO4 stock is 10 mM, which is 10,000x 1 µM

MS medium

MS medium kdorfman Thu, 05/23/2013 - 16:02

1/2 MS

1/2 MS kdorfman Wed, 12/22/2021 - 20:25

*Add the other salt mixtures to the water to prevent precipitation

**pH to 5.7 with 1M KOH (initial pH = ~3.66) (needs~21 drops or ~720 µL/L)

Autoclave with stir bar in flask or bottle

Stir until cool enough to handle

Pour 30 mL per plate (use the deep ones)

Pour 50 mL per square plate (20 plates per liter)

[1]: Sigma M 0654 Murashige and Skoog basal salt macronutrient solution (Krackeler 45-M0654-1L-EA) ~$26

[2]: Sigma M 0529 Murashige and Skoog basal salt micronutrient solution (Krackeler 45-M0529-1L-EA) ~$26

1X MS

1X MS kdorfman Wed, 12/22/2021 - 20:37

*Add the other salt mixtures to the water to prevent precipitation

**pH to 5.7 with 1M KOH (initial pH = ~3.66) (needs~21 drops or ~720 µL/L)

Autoclave with stir bar in flask or bottle

Stir until cool enough to handle

Pour 30 mL per plate (use the deep ones)

[1]: Sigma M 0654 Murashige and Skoog basal salt macronutrient solution (Krackeler 45-M0654-1L-EA) ~$26

[2]: Sigma M 0529 Murashige and Skoog basal salt micronutrient solution (Krackeler 45-M0529-1L-EA) ~$26

To make	plates low-iron 1/2 MS
start with	mL ddH2O (~ 60% final volume)
add:	mL 10X macronutrients
add:	mL boric acid 1000X
add:	mL cobalt chloride 10,000X
add:	mL cupric sulfate 10,000X
add:	mL ferrous sulfate 10 mM
add:	mL KI 10,000X
add:	mL manganese sulfate 1000X
add:	mL moybdic acid 10,000X
add:	mL zinc sulfate 1000X
add:	g MES, pH to 5.7 with KOH
bring to	mL final volume
add:	g bacto- or phyto-agar
autoclave for	minutes

To make	mL LPGM
at	% sucrose
add:	mL 150 mM MES
add:	uL 10 mM CaCl2
add:	uL 160 mM Borate, pH to 5.7 with KOH
add:	g sucrose

To make	plates low-iron MS
start with	mL ddH2O (~ 60% final volume)
add:	mL 10X macronutrients
add:	mL boric acid 1000X
add:	mL cobalt chloride 10,000X
add:	mL cupric sulfate 10,000X
add:	mL KI 10,000X
add:	mL manganese sulfate 1000X
add:	mL moybdic acid 10,000X
add:	mL zinc sulfate 1000X
add:	g MES, pH to 5.7 with KOH
bring to	mL final volume
add:	g bacto- or phyto-agar
autoclave for	minutes

To make	plates low-iron MS
start with	mL ddH2O (~ 60% final volume)
add:	mL 10X macronutrients
add:	mL boric acid 1000X
add:	mL cobalt chloride 10,000X
add:	mL cupric sulfate 10,000X
add:	mL ferrous sulfate 10 mM
add:	mL KI 10,000X
add:	mL manganese sulfate 1000X
add:	mL moybdic acid 10,000X
add:	mL zinc sulfate 1000X
add:	g MES, pH to 5.7 with KOH
bring to	mL final volume
add:	g bacto- or phyto-agar
autoclave for	minutes

To make	30 mL plates
start with:	mL water initial volume*
add:	mL 10X macronutrients [1]
add:	mL 10X micronutrients [2]
add:	g MES. pH to 5.7 w/ 1M KOH**
bring volume to:	mL water final volume
add:	g bacto or phyto agar
autoclave for:	minutes

Plant growth media

1/2 MS low Iron

LPGM

MS high salt

MS iron-free

MS low iron

MS medium

1/2 MS

1X MS