SC -Leu High Ade

SC -Leu High Ade kdorfman Sun, 01/19/2020 - 16:39

Plates for Jeff Laney's Cell & Molecular Biology Lab

8X SC-HLT hi Ade concentrate

8X SC-HLT hi Ade concentrate kdorfman Fri, 01/22/2021 - 22:01

Concentrate to make SC-HLT High Ade agar

add 125 mL heated concentrate to autoclaved 875 mL water + 20 g agar right out of the autoclave and still hot

to make enough concentrate for:
Liter(s) SC-HLT high Ade agar medium
start with
mL water
add:
g YNB
and:
g ammonium sulfate
and:
g SC -HLT
autoclave for:
minutes (to get it into solution)
while it's still hot, add:
g glucose
and:
g Adenine hemi sulfate
and:
g tryptophane
and:
mL his 100X concentrate.
Bring to a final volume of:
mL 8X SC-L high Ade and filter sterilize

Laney's SC high ade

Laney's SC high ade kdorfman Sun, 01/26/2020 - 16:22

From Jeff Laney's recipe:

Per liter of medium, mix the following asceptically, and pre heat:

Component Volume (mL)
10X SC-AHLT 100
10X YNB+AS 100
10X 20% glucose 100
10X Adenine hemi sulfate 100
50X Tryptophan 20
100X Histidine HCl 10

Add to 20g agar in 667.5 mL water, autoclaved 30 minutes

Mark with one green line.

SC -Leu High Ade 4x + powder

SC -Leu High Ade 4x + powder kdorfman Sun, 01/26/2020 - 16:26

SC -Leu High Ade 4x + powder

Use the 4x SC-AHLT to mix dry ingredients

(4x already made; not enough dry left to start over.)

per Liter:

Agar 20 g + 620 mL water. Autoclave

Meanwhile, make:

Component Volume (mL)
4X SC-AHLT 250
10X 20% glucose 100
50X Tryptophan 20
100X Histidine HCl 10

Add dry:

Component g
YNB 1.7
Adenine hemi sulfate 0.2
Ammonium Sulfate 5

Filter sterilize, warm up, then add to molten agar

Mark with one green line

SC -Leu High Ade 4X

SC -Leu High Ade 4X kdorfman Sun, 01/26/2020 - 16:23

Using dilution factor 4:
(Because 10x won't go into solution)

But the agar is so viscous that bubbles don't reach the surface. Hard to pour.

To make
SC-leu high ade agar plates
with mm plate diameter
pour mL per plate
start with: mL water
add: g agar, leave stir bar in
autoclave for: minutes
Meanwhile, using X SC-AHLT
Asceptically mix together,
warm up, then add to autoclaved agar:		 mL SC-AHLT,
and : mL 10x YNB+AS ,
and: mL 20% glucose ,
and: mL 10X Adenine hemi sulfate,
and: mL 50X Tryptophan,
and: mL 100X Histidine HCl.
for a final volume of mL

Mark with one green line

SC-L high Ade from concentrate

SC-L high Ade from concentrate kdorfman Fri, 01/22/2021 - 20:55
To make SC -L high Ade plates (100 mm)
mix: g agar
into: mL water
autoclave for: minutes
while it's still hot, asceptically add: mL heated 8X SC-L high Ade concentrate
for a final volume of: mL. Mix thoroughly before pouring plates

Mark with one green line.

SC-L high Ade mostly powder

SC-L high Ade mostly powder kdorfman Tue, 02/04/2020 - 13:41

High concentration agar is difficult to work with.

  • Tends to boil over, so needs a lot of water in the autoclave pan
  • Hard to get into solution - frequently needs re-autoclaving
  • So viscous that bubbles don't rise to the surface

So

  • mix the ingredients that can't be autoclaved in as little water as possible
  • Only use high concentration stock solutions for micro-ingredients

Per Liter:

dry ingredient g
SC-AHLT 1.64
YNB 1.7
Ammonium sulfate 5
glucose 20
stock solutions mL
50X Tryptophan 20
100X Histidine HCl 10
10X Adenine hemi sulfate 100

plus water to 150 mL

filter sterilize

Autoclave

  • 20 g agar
  • 850 mL water

Mix and pour