Mark,
Is there any concern about breaking tubes by tightening the caps after the autoclave? IE vacuum pressure caused by sealing caps while still hot.
I made myself a step by step list. Would you mind checking to see if I missed something? Bare in mind I'll be using pre-sterilized plastic plates.
Yeast Banking
Prep:
Make 100ml of 5% w/v starter wort
5g DME 100ml DI water, stirplate 2 min, microwave to boil
Make 500ml of 5% w/v and 1.5% w/v agar medium
25g DME, 1.5g agar, 500ml DI water, stirplate 2 min, microwave to boil
Pour agar medium into 5 slants 1/2 full, cap on lose
Pour starter wort into 3 slants 1/2 full, cap on lose
Place all 8 slants and agar vessel into autoclave 15min at 15psi
When done and cooled to 150º tighten slant caps and pour 4 plates near alcohol lamp
Lay slants at angle to cool, once cool store upside down to keep condensation away from agar
Making storage slants:
Work near alcohol lamp, flaming loop, openings and caps
Gather sample from commercial sample or old slant, streak one plate, incubate 24-48 hrs
Select a single colony and inoculate a slant, repeat for a second slant, incubate till mostly covered
Store upside down in zip lock bag in fridge up to 6 months
Making a starter
Work near alcohol lamp, flaming loop, openings and caps
Collect a sample from best slant, streak two plates, incubate at room temp 24-48 hrs
Dispose or clean old slants
Select single colony and inoculate tube of starter wort, repeat for second tube of starter
Select single colony and inoculate new storage slant, repeat for second storage slant.
Incubate 48 hrs, store slants in zip lock bag in fridge
Step up tube starter to 100ml starters, then to 500ml starters, then decant and pitch to 1LO2HK starter
Timing:
Slant a yeast sample if not brewing for a month or more, otherwise just step start and refrigerate the 500ml till brew day
Store slants up to 6 months before re-slanting
Plate out a slant sample for step starters 8 days before brewday
2 days to incubate plate
2 days to incubate 10ml starter
2 days to incubate 100ml starter
2 days to incubate 500ml starter
Morning of brew day decant and pitch to 1LO2HK Starter