Team:UNICAMP-EMSE Brazil/Notebook/16 September 2011

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May
MTWTFSS
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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
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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
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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
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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, 16/09/2011.
Gel 1: pSB1/ pSB 2/ pSB 3/ pSB 4/ pSB 5/ SoxS+ RBS+HlyB +RBS+HlyD+ RBS+TolC+T C1 1/ SoxS+ RBS+HlyB+ RBS+HlyD + RBS+TolC+T C1 2/ SoxS+ RBS+HlyB+ RBS+HlyD + RBS+TolC+T C1 3/ RBS+IL-12 C4/ RBS+IL-12 C5/ RBS+HlyD C4/ RBS+HlyD C5
Electrophoresis Gel 2, 16/09/2011.
Gel 2: RBS+Tol C C10/ RBS+Tol C C11/ RBS+QseB+RBS+QseC C10/ RBS+QseB+RBS+QseC C10/ RBS+QseB+RBS+QseC C11/ RBS+QseB+RBS+QseC C12/ RBS+SoxS C10/ RBS+SoxS C11/ RBS+SoxS C12/ RBS+SoxS C13/ RBS+HlyB C10/ RBS+HlyB C11/ RBS+HlyB C12/ (E-S)

16 September 2011

Isolate and cut inserts

Objective:

  1. Quantification of linearized pSB1C3 vector purified from the gel (volume of ladder applied 6 uL; sample: 5 uL DNA +1 uL buffer),
  2. Checking digestion ( EcoRI and PstI) of samples mini-preped 09/15 and 09/16.
  3. Checking digestion of SoxS+HlyB+HlyD +TolC+T C1 1/ SoxS+HlyB+HlyD +TolC+T C1 2/ SoxS+HlyB+HlyD +TolC+T C1 3 (EcoRI and PstI) of samples mini-preped 09/08.

Samples:

  • Gel 1:
  • pSB1/ pSB 2/ pSB 3/ pSB 4/ pSB 5/ SoxS+ RBS+HlyB +RBS+HlyD+ RBS+TolC+T C1 1/ SoxS+ RBS+HlyB+ RBS+HlyD + RBS+TolC+T C1 2/ SoxS+ RBS+HlyB+ RBS+HlyD + RBS+TolC+T C1 3/ RBS+IL-12 C4/ RBS+IL-12 C5/ RBS+HlyD C4/ RBS+HlyD C5
  • Gel 2:
  • RBS+Tol C C10/ RBS+Tol C C11/ RBS+QseB+RBS+QseC C10/ RBS+QseB+RBS+QseC C10/ RBS+QseB+RBS+QseC C11/ RBS+QseB+RBS+QseC C12/ RBS+SoxS C10/ RBS+SoxS C11/ RBS+SoxS C12/ RBS+SoxS C13/ RBS+HlyB C10/ RBS+HlyB C11/ RBS+HlyB C12/
  • Ladder:
  • 100-10.000 bp (Fermentas)
  • Gel Agarose concentration:
  • 1,0%


  • Results:

Construction Total size (gene+vector) Linear vector size Gene size Result
pSB 1 2070 2070 ---- Small concentration
pSB 2 2070 2070 ---- Small concentration
pSB 3 2070 2070 ---- Small concentration
pSB 4 2070 2070 ---- Small concentration
pSB 5 2070 2070 ---- Small concentration
SoxS+ RBS+HlyB+ RBS+HlyD + RBS+TolC+T C1 1 8662 3205 5457 Digestion did not work
SoxS+ RBS+HlyB+ RBS+HlyD + RBS+TolC+T C1 2 8662 3205 5457 Digestion did not work
SoxS+ RBS+HlyB+ RBS+HlyD + RBS+TolC+T C1 3 8662 3205 5457 Strange band pattern
RBS+IL-12 C4 3728 2070 1658 OK
RBS+IL-12 C5 3728 2070 1658 Probably digestion did not work
RBS+HlyD C4 3566 2070 1496 Strange band pattern. Can it be HlyB instead of HlyD?
RBS+HlyD C5 3566 2070 1496 No fragment visible band
RBS+Tol C C10 3650 2070 1580 Strange band pattern
RBS+Tol C C11 3650 2070 1580 Strange band pattern
RBS+QseB+RBS+QseC C10 4156 2070 2086 Has two bands that matches correct sizes, but an extra strange band
RBS+QseB+RBS+QseC C11 4156 2070 2086 Strange band
RBS+QseB+RBS+QseC C12 4156 2070 2086 Has two bands that matches correct sizes, but an extra strange band
RBS+SoxS C10 2173 2070 103 Seems little bigger than expected
RBS+SoxS C11 2173 2070 103 Seems little bigger than expected
RBS+SoxS C12 2173 2070 103 Seems little bigger than expected
RBS+SoxS C13 2173 2070 103 Seems little bigger than expected
RBS+HlyB C10 4253 2070 2183 Non match band size. Where´s the vector band?
RBS+HlyB C11 4253 2070 2183 Non match band pattern. Cain it be HlyD?
RBS+HlyB C12 4253 2070 2183 Non match band size. Where´s the vector band?