Team:UNICAMP-EMSE Brazil/Notebook/29 August 2011

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Contents

Notebook

Click on a date to see what we have done!

May
MTWTFSS
            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 31
June
MTWTFSS
    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
MTWTFSS
        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 31
August
MTWTFSS
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 31
September
MTWTFSS
      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
October
MTWTFSS
          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
31


Electrophoresis Gel 1, 29/08/2011.
Gel 1: HlyA+Terminator C1/ HlyA+Terminator C2/ HlyA+Terminator C3/ HlyA+Terminator C4/ Const. Promoter+ RBS+SoxR+ Terminator C1/ Const. Promoter+ RBS+SoxR+ Terminator C2/ SoxS+RBS+IL 10 C1/ SoxS+RBS+IL 10 C2/ flHDC+IL 12 C1/ flHDC+IL 12 C2/ RBS+HlyD+TolC+Terminator C2a/ RBS+HlyD+TolC+Terminator C1a
Electrophoresis Gel 2, 29/08/2011.
Gel 2: SoxS+RBS+GFP C1/ SoxS+RBS+GFP C2/ RBS+HlyD+TolC+Terminator C1/ RBS+HlyD+TolC+Terminator C2/ flHDC+RBS+HlyB C1/ flHDC+RBS+HlyB C2/ SoxS+RBS+HlyB C1/ SoxS+RBS+HlyB C2/ flHDC+RBS+GFP C1/ flHDC+RBS+GFP C2/ Constitutive Promoter+ RBS+QseB+RBS+QseC+Terminator C1/ Const. Promoter+ RBS+QseB+RBS+QseC+Terminator C2/ Const. Promoter+ RBS+QseB+RBS+QseC+Terminator C3/ Const. Promoter+ RBS+QseB+RBS+QseC+Terminator C4.

29 August 2011

Finishing the first two biobricks!!!

Objective:

  • Checking digestion of yesterday's ligation with EcoRI and PstI

Samples:

  • Gel 1:
  • HlyA+Terminator C1/ HlyA+Terminator C2/ HlyA+Terminator C3/ HlyA+Terminator C4/ Const. Promoter+ RBS+SoxR+ Terminator C1/ Const. Promoter+ RBS+SoxR+ Terminator C2/ SoxS+RBS+IL 10 C1/ SoxS+RBS+IL 10 C2/ flHDC+IL 12 C1/ flHDC+IL 12 C2/ RBS+HlyD+TolC+Terminator C2a/ RBS+HlyD+TolC+Terminator C1a
  • Gel 2:
  • SoxS+RBS+GFP C1/ SoxS+RBS+GFP C2/ RBS+HlyD+TolC+Terminator C1/ RBS+HlyD+TolC+Terminator C2/ flHDC+RBS+HlyB C1/ flHDC+RBS+HlyB C2/ SoxS+RBS+HlyB C1/ SoxS+RBS+HlyB C2/ flHDC+RBS+GFP C1/ flHDC+RBS+GFP C2/ Constitutive Promoter+ RBS+QseB+RBS+QseC+Terminator C1/ Const. Promoter+ RBS+QseB+RBS+QseC+Terminator C2/ Const. Promoter+ RBS+QseB+RBS+QseC+Terminator C3/ Const. Promoter+ RBS+QseB+RBS+QseC+Terminator C4
  • Ladder:
  • 100-10.000 bp (Fermentas)
  • Gel Agarose concentration:
  • 1,5%


  • Results:

Gene Total size (gene+vector) Linear vector size Gene size Result
HlyA+Terminator C1 3527 bp 3205 bp 322 bp No visible band
HlyA+Terminator C2 3527 bp 3205 bp 322 bp OK
HlyA+Terminator C3 3527 bp 3205 bp 322 bp OK
HlyA+Terminator C4 3527 bp 3205 bp 322 bp OK
Const. Promoter+ RBS+SoxR+ Terminator C1 2732 bp 2079 bp 653 bp Seems smaller
Const. Promoter+ RBS+SoxR+ Terminator C2 2732 bp 2079 bp 653 bp Seems smaller
SoxS+RBS+IL 10 C1 3543 bp 2800 bp 743 bp No visible bands
SoxS+RBS+IL 10 C2 3543 bp 2800 bp 743 bp OK
flHDC+IL 12 C1 4584 bp 2800 pb 1784 No visible bands
flHDC+IL 12 C2 4584 bp 2800 bp 1784 OK
RBS+HlyD+TolC+Terminator C2a 6376 bp 3205 bp 3171 bp Run more
RBS+HlyD+TolC+Terminator C1a 6376 bp 3205 bp 3171 bp Run more
SoxS+RBS+GFP C1 3638 bp 2800 bp 838 bp No visible band
SoxS+RBS+GFP C2 3638 bp 2800 bp 838 bp OK
RBS+HlyD+TolC+Terminator C1 6376 bp 3205 bp 3171 bp Run more
RBS+HlyD+TolC+Terminator C2 6376 bp 3205 bp 3171 bp Run more
flHDC+RBS+HlyB C1 5109 bp 2800 bp 2309 bp OK
flHDC+RBS+HlyB C2 5109 bp 2800 bp 2309 bp No visible band
SoxS+RBS+HlyB C1 5086 bp 2800 bp 2286 bp OK
SoxS+RBS+HlyB C2 5086 bp 2800 bp 2286 bp OK
flHDC+RBS+GFP C1 3661 bp 2800 bp 861 bp No visible bands
flHDC+RBS+GFP C2 3661 bp 2800 bp 861 bp No visible bands
Constitutive Promoter+RBS+
QseB+RBS+QseC+Terminator C1
4295 bp 2079 bp 2216 bp Run more
Constitutive Promoter+RBS+
QseB+RBS+QseC+Terminator C2
4295 bp 2079 bp 2216 bp Run more
Constitutive Promoter+RBS+
QseB+RBS+QseC+Terminator C3
4295 bp 2079 bp 2216 bp Run more
Constitutive Promoter+RBS+
QseB+RBS+QseC+Terminator C4
4295 bp 2079 bp 2216 bp Run more




Electrophoresis Gel 3, 29/08/2011.
Gel 1: HlyA+Terminator C1/ HlyA+Terminator C2/ HlyA+Terminator C3/ HlyA+Terminator C4/ Const. Promoter+ RBS+SoxR+ Terminator C1/ Const. Promoter+ RBS+SoxR+ Terminator C2/ SoxS+RBS+IL 10 C1/ SoxS+RBS+IL 10 C2/ flHDC+IL 12 C1/ flHDC+IL 12 C2/ RBS+HlyD+TolC+Terminator C2a/ RBS+HlyD+TolC+Terminator C1a
Electrophoresis Gel 4, 29/08/2011.
Gel 4: SoxS+RBS+GFP C1/ SoxS+RBS+GFP C2/ RBS+HlyD+TolC+Terminator C1/ RBS+HlyD+TolC+Terminator C2/ flHDC+RBS+HlyB C1/ flHDC+RBS+HlyB C2/ SoxS+RBS+HlyB C1/ SoxS+RBS+HlyB C2/ flHDC+RBS+GFP C1/ flHDC+RBS+GFP C2/ Constitutive Promoter+ RBS+QseB+RBS+QseC+Terminator C1/ Const. Promoter+ RBS+QseB+RBS+QseC+Terminator C2/ Const. Promoter+ RBS+QseB+RBS+QseC+Terminator C3/ Const. Promoter+ RBS+QseB+RBS+QseC+Terminator C4.

Same electrophoresys but after a longer period run:

Samples:

  • Gel 3:
  • HlyA+Terminator C1/ HlyA+Terminator C2/ HlyA+Terminator C3/ HlyA+Terminator C4/ Const. Promoter+ RBS+SoxR+ Terminator C1/ Const. Promoter+ RBS+SoxR+ Terminator C2/ SoxS+RBS+IL 10 C1/ SoxS+RBS+IL 10 C2/ flHDC+IL 12 C1/ flHDC+IL 12 C2/ RBS+HlyD+TolC+Terminator C2a/ RBS+HlyD+TolC+Terminator C1a
  • Gel 4:
  • SoxS+RBS+GFP C1/ SoxS+RBS+GFP C2/ RBS+HlyD+TolC+Terminator C1/ RBS+HlyD+TolC+Terminator C2/ flHDC+RBS+HlyB C1/ flHDC+RBS+HlyB C2/ SoxS+RBS+HlyB C1/ SoxS+RBS+HlyB C2/ flHDC+RBS+GFP C1/ flHDC+RBS+GFP C2/ Constitutive Promoter+ RBS+QseB+RBS+QseC+Terminator C1/ Const. Promoter+ RBS+QseB+RBS+QseC+Terminator C2/ Const. Promoter+ RBS+QseB+RBS+QseC+Terminator C3/ Const. Promoter+ RBS+QseB+RBS+QseC+Terminator C4
  • Ladder:
  • 100-10.000 bp (Fermentas)
  • Gel Agarose concentration:
  • 1,5%



  • Results:

Gene Total size (gene+vector) Linear vector size Gene size Result
RBS+HlyD+TolC+Terminator C2a 6376 pb 3205 pb 3171 pb Superposition of the bands,
but wont be a problem in the
next ligation, we will linearize only
RBS+HlyD+TolC+Terminator C1a 6376 pb 3205 pb 3171 pb Superposition of the bands,
but wont be a problem in the
next ligation, we will linearize only
RBS+HlyD+TolC+Terminator C1 6376 pb 3205 pb 3171 pb Superposition of the bands,
but wont be a problem in the
next ligation, we will linearize only
RBS+HlyD+TolC+Terminator C2 6376 pb 3205 pb 3171 pb Superposition of the bands,
but wont be a problem in the
next ligation, we will linearize only
Constitutive Promoter+ RBS+QseB+RBS+QseC+Terminator C1 4295 pb 2079 pb 2216 pb Seems to be OK
Const. Promoter+ RBS+QseB+RBS+QseC+Terminator C2 4295 pb 2079 pb 2216 pb Seems to be OK
Const. Promoter+ RBS+QseB+RBS+QseC+Terminator C3 4295 pb 2079 pb 2216 pb Seems to be OK
Const. Promoter+ RBS+QseB+RBS+QseC+Terminator C4 4295 pb 2079 pb 2216 pb Seems to be OK



OBS:-Inoculation of new colonies of flHDC+RBS+IL-12 and flHDC+RBS+GFP


Wowwww first two biobricks assembled !!! :) S2

UNICAMP-EMSE Brazil Biobrick1 29 08 11.png UNICAMP-EMSE Brazil Biobrick2 29 08 11.png