Team:Kent/Notebook


iGEM Kent 2015

Notebook

June

M T W T F S S
1 2 3 4 5 6 7
8 9 10 11 12 13 14
15 16 17 18 19 20 21
22 23 24 25 26 27 28
29 30

July

M T W T F S S
1 2 3 4 5
6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 2930 31

August

M T W T F S S
1 2
3 4 5 6 7 8 9
10 11 12 13 14 15 16
17 18 19 20 21 22 23
24 25 2627 28 29 30
31

September

M T W T F S S
1 2 3 4 5 6
7 8 9 10 11 12 13
14 15 16 17 18 19 20
21 22 23 24 25 26 27
28 29 30



















Day 1 Wet lab 22/06/15

  • Made LB plates
  • Made LB broth
  • TIPS autoclaved
  • Day 2 Wet lab 23/06/15

  • Set up Top10 overnight, VS45 overnight, US348 overnight
  • Meeting with advisors to discuss progress
  • Day 3 Wet lab 24/06/15

  • Filter sterilise the buffer
  • 250ml no antibiotic broth, put top10 cells in
  • Incubate until optical density is 0.6
  • Miniprep PSB1C3 plasmid
  • Day 4 Wet lab 25/06/15

  • Transformation
  • Prepare SBC media
  • Digest of PSB1c3 from MS348
  • Day 5 Wet lab 26/06/15

  • Calculated competent cell efficiency
  • Agarose gel formation
  • Gel electrophoresis
  • Day 6 Wet lab 29/06/15

    Transforming linear PSB1C3

    Day 7 Wet lab 30/06/15

  • Miniprep of PSB1A3 plasmid
  • Gel electrophoresis of a large quantity of PSB1C3
  • Meeting with advisors to discuss progress
  • Day 8 Wet lab 01/07/15

  • Gel electrophoresis of the purified DNA extracted
  • Transformation of pSB1A3 circular plasmid to VS45 cells (competent)
  • Digest of all pSB1C3 plasmids
  • Day 9 Wet lab 02/07/15

  • Competent cells transformation efficiency
  • Mini-prep of 1MS340 to get pSB1C3 plasmid
  • Gel extraction of big digest
  • Quantification of the digest. Added sample buffer to each digest
  • Day 10 Wet lab 03/07/15

  • Transformation efficiency
  • Day 11 Wet lab 06/07/15

  • TOP10 cells containing pSB1A3 with limonene synthase were cultured overnight on AMP plates
  • Colonies of VS45 containing pSB1A3 were patched onto chloramphenicol plates
  • Overnight digest of miniprepped pSBIC3 using ECORI and PSTl
  • Day 12 Wet lab 07/07/15

  • An agarose gel of the overnight digest was ran, but no bands were visible
  • Set up overnight cultures of pSBIC3 and pSBIA3 to be miniprepped the next day
  • Set up an overnight digest of pSBIC3
  • Day 13 Wet lab 08/07/15

  • Agarose gel of overnight digest was ran
  • miniprep of pSBIA3 and pSBIC3
  • Day 14 Wet lab 09/07/15

  • Agarose gel of digested pSBIA3 and pSBIC3 (no bands visible)
  • Overnight cultures of MS349 containing pSBIA3 LIMS and MS340 containing pSBIC3 were set up, ready to be miniprepped
  • Day 15 Wet lab 10/07/15

  • Miniprep of plasmids pSBIA3 and pSBIC3
  • Day 16 Wet lab 13/07/15

  • Overnight cultures of MS349 containing pSBIA3 LIMS and MS340 containing pSBIC3 were set up, ready to be miniprepped
  • Overnight digest of miniprepped plasmids pSBIA3 and pSBIC3 using the enzymes ECORI and PSTI
  • Day 17 Wet lab 14/07/15

  • Agarose gel of digested plasmids pSBIA3 and pSBIC3 - it finally worked!!!
  • Miniprep of overnight cultures
  • Gel extraction of both plasmids
  • Analytical gel using Hyperladder I to quantify the plasmid