Paris/July 20

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Contents

PCRs

PCR : Lox71-KmR-Lox66
PCR Settings Buffer (5x) 5x 10µL Expected size
Annealing (°C) MgCl2 10µM 10µM 0µL 1084
55 dNTP 10µM 10µM 1µL Success
Time Elongation Oligo F 10µM 10 Lox71-KmR-F 2.5µL  ?
3m00' Oligo R 10µM 11 Lox66-KmR-R 2.5µL Image (click to enlarge)
Number cycles Water 34µl [[Image:|30px]]
35 Polymerase Phusion 0.5µL Band (0=ladder)
DNA MP3_pSB1AK3-BBa_B0015 0.5µl
PCR : Lox71-FtsA-FtsZ
PCR Settings Buffer (5x) 5x 10µL Expected size
Annealing (°C) MgCl2 10µM 10µM 0µL 2588
55 dNTP 10µM 10µM 1µL Success
Time Elongation Oligo F 10µM 3 Lox71-FtsA-F 2.5µL  ? The PCR seemed to have worked but the purification failed
3m00' Oligo R 10µM 2 FtsZ-R 2.5µL Image (click to enlarge)
Number cycles Water 34µl [[Image:|30px]]
35 Polymerase Phusion 0.5µL Band (0=ladder)
DNA toothpick in MG1655 glycerol

Glycerol stock

  • BBa_pJ23107 clone 3&4 (S14)
  • BBa_I0500 clone 1&2 (S13)

Minipreps on Biobricks

  • BBa_pJ23107 clone 3&4
  • BBa_I0500 clone 1&2

E.Coli pKS::DGAT

We look under microscopy 36 hours after incubation of E.coli transformed by pKS::DGAT and the control E.coli transformed by part B0015 on different LB medium (See July 18).

  • Observations:

We can observe colonies (X10).

Coli dgat 07202007.jpg
  • Interpretation

We can observe lipid inclusions within colonies. Further investigations need single cell visualisation.