Team:UNICAMP-EMSE Brazil/Notebook/9 September 2011

From 2011.igem.org

Click here to see our PROTOCOLS

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, 09/09/2011.
Gel 1: SoxS+RBS+GFP+HlyA+T nd/ SoxS+RBS+GFP+HlyA+T E-P/ Const. Promoter+ RBS+SoxR+ T nd/ Const. Promoter+ RBS+SoxR+ T E-P/ SoxS+RBS+HlyB+HlyD+TolC + T nd/ SoxS+RBS+HlyB+HlyD+TolC + T E-P
Electrophoresis Gel 2, 09/09/2011.
Gel 2: SoxS+RBS+GFP+HlyA+T E-X/ / Const. Promoter+ RBS+SoxR+ T E-S

09 September 2011

Digestion and Electrophoresis

  • Digestion recipe:
  • DNA – 7ul
  • Buffer O – 2ul
  • EcoRI – 0,5ul
  • PstI – 0,5ul
  • Water – 10ul

Samples:

  • Gel 1:
  • SoxS+RBS+GFP+HlyA+T nd/ SoxS+RBS+GFP+HlyA+T E-P/ Const. Promoter+ RBS+SoxR+ T nd/ Const. Promoter+ RBS+SoxR+ T E-P/ SoxS+RBS+HlyB+HlyD+TolC + T nd/ SoxS+RBS+HlyB+HlyD+TolC + T E-P
  • Ladder:
  • 100-10.000 bp (Fermentas)
  • Gel Agarose concentration:
  • 1,0%


  • Results:

Construction Total size (gene+vector) (bp) Linear vector size (bp) Gene size (bp) Result
1) SoxS+RBS+GFP+HlyA+T 4365 3205 1160 ok
2) Const. Promoter+ RBS+SoxR+ T 2732 2079 653 ok
3) SoxS+RBS+HlyB+HlyD+TolC + T 8257 2800 5457  ??

Samples:

  • Gel 2:
  • SoxS+RBS+GFP+HlyA+T E-X/ / Const. Promoter+ RBS+SoxR+ T E-S


Check up electrophoresis

File:UNICAMP-EMSE Brazil Gel3 09 09 11.jpg
Electrophoresis Gel 3, 09/09/2011.
Gel 2: Sox S + RBS + IL10 ; flHDC + RBS +IL12+ HlyA + T (digest by (E-P))

Objective:

  • Checking digestion

Samples:

  • Gel 3:
  • Sox S + RBS + IL10 ; flHDC + RBS +IL12+ HlyA + T (digest by (E-P))
  • Ladder:
  • 100-10.000 bp (Fermentas)
  • Gel Agarose concentration:
  • 1,0%


  • Results:

Construction Total size (gene+vector) (bp) Linear vector size (bp) Gene size (bp) Result
Sox S + RBS + IL10 3543 2800 743 No Gene size band
Sox S + RBS + IL10 3543 2800 743 No Gene size band
Sox S + RBS + IL10 3543 2800 743 No Gene size band; and partial digestion
flHDC +RBS +IL12+ HlyA +T 5311 3205 2106 Ok
flHDC +RBS +IL12+ HlyA +T 5311 3205 2106 Ok