Finally Bio-Lab activity began!

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June, 3rd9° Meeting - Discussion of final and technical details about The Project. Planning of Bio-Lab activity.
June, 7th1° Bio-Lab - <partinfo>BBa_E2050</partinfo>, <partinfo>BBa_K165018</partinfo>, <partinfo>BBa_K165037</partinfo>, <partinfo>BBa_J61001</partinfo>, <partinfo>BBa_K081008</partinfo> and <partinfo>BBa_K125500</partinfo> were resuspended from Spring 2010 DNA distribution. <partinfo>BBa_P1004</partinfo> was resuspended from Spring 2009 DNA distribution. All BioBricks were transformed in E. coli DH5alpha.
Liquid LB+Amp and LB+Amp agar plates were prepared according with "protocols".
June, 8th2° Bio-Lab - Inoculum from plates in liquid Lb and glycerol stocks preparation. Falcon were re-filled for tomorrow mini-prep.
June, 9th3° Bio-Lab - MiniPrep of <partinfo>BBa_E2050</partinfo>, <partinfo>BBa_K165018</partinfo>, <partinfo>BBa_K165037</partinfo>, <partinfo>BBa_J61001</partinfo>, <partinfo>BBa_K081008</partinfo> and <partinfo>BBa_K125500</partinfo>. Digestion, gel run/cut and ligation of:
  • I0: <partinfo>BBa_K125500</partinfo> (E-S)+<partinfo>BBa_B0015</partinfo> (E-X)
  • I1: <partinfo>BBa_E2050</partinfo> (E-S)+<partinfo>BBa_K165018</partinfo> (E-X)
  • I2: <partinfo>BBa_P1004</partinfo> (E-S)+<partinfo>BBa_B0015</partinfo> (E-X)
  • I3: <partinfo>BBa_K081008</partinfo> (E-S)+<partinfo>BBa_B0015</partinfo> (E-X)
June, 10th4° Bio-Lab - Transformation of ligations I0, I1, I2, I3 in home made E. coli DH5-alpha competent cells. Transformants were plated on LB+Amp agar plates.
June, 11th5° Bio-Lab - Inoculum from plates in liquid LB and glycerol stocks preparation for I0, I1, I2 and I3 in duplicate (2 colonies picked from each plate).
June, 12thiGEM EU workshop.
June, 13thiGEM EU workshop.
June, 14th6° Bio-Lab - Inoculum from glycerol stock of I0-1, I0-2, I1-1, i1-2, I2-1, I3-1 and I3-2 for screening.
June, 15th7° Bio-Lab - MiniPrep, gel run/cut and ligation of
  • I4: <partinfo>BBa_K165037</partinfo> (S-P) + I1 (X-P)
  • I5: I2 (E-S) + <partinfo>BBa_J61001</partinfo> (E-X)
  • I6: I3 (E-S) + <partinfo>BBa_T9002</partinfo> (E-X)
June, 16th8° Bio-Lab - Tranformation of I4 (DH5-alpha), I5 (TOP10) and I6 (DH5-alpha).
June, 17th9° Bio-Lab - Colonies were picked from each plate, I4-1, I4-2, I5-1, I5-2, I6-1, I6-2 and I6-3 were grown in 1ml liquid culture and glycerol stocked (not I5-2, because no growth was observed in LB+Cm and not for I6-1, not grown). Cultures refilled with 5ml LB+antibiotic for tomorrow MiniPrep and screening.
June, 18th10° Bio-Lab - MiniPrep for I4-1, I4-2, I5-1, I6-2 and I6-3. Screening gel (all the clones are ok!) :)
June, 21st11° Bio-Lab - Week activities planning and freezer sorting.
June, 22nd12° Bio-Lab - Inoculations of I6, BBa_J23118, BBa_J23110, BBa_J23114, BBa_J23116.
June, 23rd13° Bio-Lab - 10° Meeting - Updating about Bio-Lab activity.
MiniPrep of ON cultures I6, BBa_J23118, BBa_J23110, BBa_J23114, BBa_J23116, with NanoDrop quantification. Digestions, gel run and extraction of the above. We received BBa_K208001, the Silver-fusion compatible BioBrick part that codes for phasin. Streaked them on two LB+Kan agar plates (named PhaP-1 and PhaP-2)
June, 24th14° Bio-Lab - Colonies on both PhaP-1 and PhaP-2. Inoculation of one colony per plate, with subsequent growth for 6hrs. Glycerol stocks and refill for tomorrow MiniPrep. Yesterday's ligations heated to inactivate ligase and transformed in TOP10 homemade competent cells.
June, 25th15° Bio-Lab - Plates all show colonies! Picked two colonies from each plate and incubated in 1ml LB+Amp. The cultures were grown for six hours then glycerol stocks were made. PhaP-1 and 2 MiniPrep and quantification by NanoDrop. DNA quantification was poor, so we decided to perform digestion screening and to repeat Miniprep next week for sequencing. Then they were digested and gel screened, both resulting positive.
June, 28th16° Bio-Lab - Inoculum from glycerol stock for I7-1, I7-2, I8-1, I8-2, I9-1, I9-2, I10-1 and I10-2 in 5ml LB+Amp. Cultures grown overnight. Inoculum of PhaP-1 and PhaP-2 in 5ml LB+Kan and ON growth. Received strains and plasmid from CGSC (Yale University). Details in the week view.
June, 29th17° Bio-Lab - From I7-1, I7-2, I8-1, I8-2, I9-1, I9-2, I10-1 and I10-2 5ul were aliquoted and diluted in 500ul LB+Amp for a preliminary TECAN test. Then cultures were MiniPrepped. Almost all cultures were digested. PhaP-1 and PhaP-2 were prepared or sequencing. All strains received from Yale Univesity were on blotting paper disks, that were placed on an LB agar plates, resuspended and then streaked. After plate streaking, disks were transferred in falcon tubes containing liquid LB.
June, 30th18° Bio-Lab - Inoculum of cultures for a TECAN test.