Plant growth media

Plant growth media kdorfman Thu, 05/23/2013 - 16:02

10 g agar per L

germination_media.xlsx

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

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

Fern Culture Media

Fern Culture Media kdorfman Thu, 01/11/2024 - 19:16

C Fern Manual

Carolina C-Fern materials

Pre-made fern medium, Carolina 156780, sold in 60 mL or 400 mL

Carolina recommends 10 mL per plate. I use 9 mL

To get gametophytes in all stages, sow fern spores in 12 60 mm dishes 4 times:

  • 1/15
  • 1/22
  • 1/29
  • 2/2 or Friday 2/3
C-Fern Manual

Fern Medium from scratch

Fern Medium from scratch kdorfman Fri, 01/12/2024 - 21:18

To make one liter
800 mL water
10X macronutrients: 100mL
200X micronutrients: 5 mL
100X Chelated Iron solution: 10 mL

pH to 6 with NaOH

Bring final volume to 1 L

Add 10 g Bacto agar (others may not work)

Autoclave 15 min

10X fern macronutrients

10X fern macronutrients kdorfman Fri, 01/12/2024 - 21:24

200X fern micronutrients

200X fern micronutrients kdorfman Fri, 01/12/2024 - 21:26

Fern medium from powder

Fern medium from powder kdorfman Wed, 01/24/2024 - 14:20

Carolina 156782

Mix contents of vial with 800 mL water

pH to 6 (NaOH)

Bring to 1 L

For single batch:

  • Add 10g Bacto Agar

  • Autoclave 45 min

For small batches:

  • Aliquot 167 mL into 6 200 mL media bottles

  • Add 1.67 g Bacto Agar to each bottle

  • Autoclave 30 min

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

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

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

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

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

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

MS medium

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

1/2 MS

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

*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
To make 30 mL plates
add: 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

*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