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Materials

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LB Media

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Auto Inducing Media

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TB Media

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LB-Miller

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ZY Media

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TB

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LB-Lennox

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20x NPS 50mL/L

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10x Salts

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LB-Luria

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50x 5052 20mL/L

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table 01 - checkbox for your media of choice

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glass

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plastic

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baffled

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unbaffled

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Equipment
Avanti JXN-26 #2, 1000mL bottles  for cell harvesting
S1 Eppendorf 5920R for cell washing with PBS
Incubator

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Innova S441 Top

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Innova S441 Mid

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Innova S441 Bottom

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Innova 4400 Top

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Innova 4400 Bottom

Preparations: - please ensure all materials needed are available in sufficient amounts
                     - please book all equipment needed online in PPMS (webinterface or QR codes)

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Time

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OD600

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Time

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OD600

- take 1mL sample for SDS PAGE analysis, freeze in -20°C
- induce protein expression by setting temperatur to 30°C/20°C
- continue growth for __________ h at __________ °C and __________ RPM overnight (e.g. 16h, 20°C, 180 RPM)

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Time

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OD600

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Mainculture Auto Inducing Media
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bevore harvesting check OD600

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Time

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OD600

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Harvesting

- transfer expression culture into 1000mL bottles for Avanti JXN-26 #2
- balance tubes correctly
- harvest cells at __________ xg for __________ min at __________ °C (e.g. 5000xg, 15min, 4°C)
- weigh 50mL falcon tubes to calculate wet cell mass

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1

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2

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3

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4

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50mL falcon tube empty

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------“------       With cells

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Wet Cell Mass

please note: supernatants will be kept in expression flasks for autoclaving, do not put them into the sink
- discard supernatant and scrape out cell pellet with soft spatula
- transfer two cell pellets into one 50mL falcon tube
- add 30mL icecold 1x PBS per 50mL falcon tube and resuspend by pipetting
- pellet cells again by centrifuging at 3234xg  for 30min at 4°C
- disard supernatant and freeze pellets with liquid Nitrogen and keep at -80°C unti further use

Results

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Liters medium

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Amount cells

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Amount cells per Liter

Create from template
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templateName157351952
templateId157351952
title@currentDate Expression in E.coli
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titleAppendix

View file
name20200205_Expression in Ecoli TEMPLATE_MACRO.docx
height250

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