IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-7-21

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To-do

  1. Inoculate liquid cultures with retransformed TP strain from yesterday-- done
  2. Extract and purify LC1PUR1 and LC2PUR2 (3kb and 400b bands) from PCR Extraction #5 of 2006-7-19.
  3. Ligate gel-purified PCR product with Topo, transform with Top10 competent cells, spread plates
  4. Wiki cleanup-- in progress
  5. Update incubator status
  6. Work on Monday presentation outline
  7. Send out signed synthesis contract with GeneArt-- done

Strike three

Once again we left a bucket of ice out on the benchtop overnight. This one contained Invitrogen's Zero-Blunt Topo cloning kit. We've labeled the box as possibly bad.

Synthesis contract

  1. Sign contract (3x)
  2. Billing - mailto address needs to be changed
  3. Add comment to ship as soon as ready

Inoculation

  • Each of the plates that were transformed from last night had lots of growth.
  • One inoculation was made for each of the plates.
  • One colony was taken from each of the plates to prevent mixes of colonies being grown up.
  • The inoculations consisted of:
    • 5 mL LB liquid culture
    • 2 um Kan
    • one colony

Gel extraction

We extracted 4 different bands from the gel that we ran yesterday:

  • LC1 3kb fragment
    • 85mg
  • LC1 400bp fragment
    • 197.5mg
  • LC2 3kb fragment
    • 132.9mg
  • LC2 400bp fragment
    • 213.3mg

Gel Purification

  • LC1 3kb fragment
    • Use 255 ul
  • LC1 400bp fragment
    • 592.5 ul
  • LC2 3kb fragment
    • 398.7 ul
  • LC2 400bp fragment
    • 639.9 ul
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