TODO: Expression Module

[bmiles]

Create module that provides general autoprotocol for performing a growth and expression of target gene.

Create a module that describes transformation of BL21 (or other expression host) with mutant plasmid.

Steps from https://github.com/dacarlin/bagel-protocol/blob/master/production.md:

Day 3: Expression cultures

In this step, you will exchange the spent growth media in your cultures with fresh growth media supplemented with isopropyl β-D-1-thiogalactopyranoside (IPTG). IPTG induces expression of your target protein. Molecular details of how the expression system works.

Material	Amount per reaction
Growth cultures from day 2	1
Terrific Broth	5 mL
IPTG solution	5 μL
Kanamycin solution	5 μL
Tube seal	1

Minirecipe: 5 mL induction medium = 5 mL Terrific Broth + 5 µL kanamycin solution + 5 µL IPTG solution

Mise en place

• tube rack
• vortexer
• P20 and tips
• serological pipet and tips

  Hardware

• centrifuge
• shaking incubator at 37 C

  Instructions

  1. Centrifuge growth cultures at 4,700 RPM for 10 minutes
  2. Peel the seals off and pour away supernatant from cell pellet
  3. Weigh the pellet and record your pellet weight
  4. Make a batch of induction media (make enough for n+1)
  5. Add 1 mL induction medium to cell pellet in tube
  6. Vortex to resuspend pellet
  7. Add 4 mL more induction medium to tube
  8. Seal with tube seals
  9. Incubate with shaking at 18 C for 24 hours

     Tips for today

• Challenge: Devise a clever way to simplify resuspension so that a robot could do it

[bmiles]

Weigh the pellet and record your pellet weight

I'm concerned there may not be a repeatable way to do this. Perhaps we can can pick colonies from a plate, growth to exponential phase then dope the plate with IPTG?

Incubate with shaking at 18 C for 24 hours

18C could be achieved with a thermocycler however container won't be compatible. For the containers we'll be growing in we can only access 37C. Is there any flexibility on the 18C @dacarlin ? If not I can look into getting 18C...

[dacarlin]

No need to weigh the pellets, A600 over time is sufficient for QC for growth. We can test other temperatures, but 18 C is what we use. The more traditional method would be to grow 12 hours at 37 C, add IPTG, then grow another 2 hours at 37 C. We could test that