Team:IISER-Pune-India/TeamNotebook/Team Notebook 11e9906b405543a29361e38a50c81024/Wet Lab Questions 2 b2c1c659471543cdbd9dfcf27808e849

Wet Lab Questions 2

Wet Lab Questions 2

Date
DepartmentWet Lab
DescriptionQuestions to answer
HP sub-branch
Links/media
Participants
Property
Property 1
Property 2
TypeDeliverables

Cyanobacteria:

1. Pick a promoter 2. Why is there a difference between yields between Lin (2020) and all other papers? 3. Effect of super-strong promoters on membrane integrity? 4. Read up more on Triparental mating, gibson method assembly, golden gate cloning, suicide vectors 5. Are there self-replicating vectors in cyanobacteria? or do genes have to be inserted into genome? 6. How can sps be overexpressed in cyanobacteria? Should we even do it? 7. Should we consider PCC11801 because it can be transformed naturally 8. Contact jayakannan for help with yields and converting one form to the other 9. Stats on cyanobacteria efficiency (particularly UTEX) 10. Contact more authors 11. Read up remaining papers and finish the table 12. Compare PCC 11801 growth rates, and ease of transformation with UTEX 2973

E coli:

1. Picking a strain 2. Plasmids used in the papers you've looked into + cloning/knock out methods they've used in paper + newer methods we could try 3. Culture conditions for succinate production and how that differs from co-culture conditions 4. E coli sucrose consumption optimisation - E coli W (with cscR knockout) vs E coli transformed with cscA, cscB, cscK 5. A priority list of gene modifications for succinate production 6. Yields of succinate production in your paper, compare it to yields in the most recent anaerobic methods, succinate yields directly from cyanobacteria 7. Succinate stats on current production methods, problems with it 8. Check out all the papers I've put up on notion that contain the keywords (E coli, Succinate, E coli + sucrose) 9. Document all the details of the papers you've looked into a table 10. Find out about the strains that we might commonly find at IISER 11. Find out about the kind of incubators we might find at IISER, do any of them have controls over CO2 input? 12. Come up with a list of specific questions you want answered by Yazdani and Amrita 13. Effect of salt on E coli, effect of succinate on pH, on E coli and Cyanobacteria growth

14. What happens due to the lack of fumarate?

Co-culture:

  1. Read up on how the co-culture is set up
  1. Should we acclimate cyanobacteria and E coli to salt?
  1. Should we allow cyanobacteria to grow and reach a stationary state before adding in the E coli?
  1. What is the procedure for encapsulation? How accessible are those hydrogel materials? How are measurements (electron microscope) done?