= MAY =
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1.ArsR-CzrA 97 E/P.
2.ArsR-CzrA 98 E/P.
3.ArsR-CzrA 99 E/P.
4.CI 02 E/P.
5.P4 PCR.
-
We extracted the correct band by kit
1.ArsR-CzrA 97 E/P.
2.ArsR-CzrA 98 E/P.
3.ArsR-CzrA 99 E/P.
4.CI 02 E/P.
5.P4 PCR.
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1. 1 kb ladder.
2.00179 Lasr lysis
.
3.00179 LasR.
4.00179 Lasr (-).
5.LasR 00179 new primers.
6.LasR 00179 new primers.
7.LasR 00179 new primers (-).
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1.1 kb ladder.
3.ArsR-CzrA 97 E/P.
5.ArsR-CzrA 98 E/P.
7.ArsR-CzrA 99 E/P.
9.CI 02 E/P.
11.AmyE 5’ ArsR-CzrA 97 E/S.
13.AmyE 5’ ArsR-CzrA 98 E/S.
15.AmyE 5’ ArsR-CzrA 99 E/S.
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1.ArsR-CzrA 97 E/P.
2.ArsR-CzRA 98 E/P.
3.1 kb ladder.
4.ArsR-CzRA 99 E/P.
5.CI 02 E/P.
6.AmyE 5’ 97 E/S.
7.AmyE 5’ 98 E/S.
8.1 kb ladder.
9.AmyE 5’ 99 E/S.
10.LasR PCR.
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1.ArsR-CzrA 97 E/P.
2.ArsR-CzRA 98 E/P.
3.1 kb ladder.
4.ArsR-CzRA 99 E/P.
5.CI 02 E/P.
6.AmyE 5’ 97 E/S.
7.AmyE 5’ 98 E/S.
8.1 kb ladder.
9.AmyE 5’ 99 E/S.
10.LasR PCR.
09/14/12
We extracted the correct band by kit GEL EXTRACTION PROTOCOL.
>We transformed ligation psB1c3+ArsR-CzrA 97/98/99/CI in competent DH5α cells from the pBad/pXyl AND team and in our own DH5α competent cells TRANSFORMATION PROTOCOL.
09/15/12
>We checked our transformations an did liquid cultures and passed the colonies to new plates LIQUID CULTURE PROTOCOL.
>We digested E0040+B0014 DIGESTIONS PROTOCOL.
>We ligated 97/98/99 psBIC3 LIGATION PROTOCOL.
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1.AmyE 5’ 97 S/P.
2.
3.AmyE 5’ 98 S/P.
4.
5.1 kb ladder.
6.AmyE 5’ 99 S/P.
7.AmyE 5’ 99 S/P.
8.E0040 B0014 X/P.
9.
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1.E0040 B0014 X/P to extract.
2.1 kb ladder.
3.Lysis.
4.-19. Amy 5’ 99/97/P4CIE1010B0014 lysis.
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1.LasR PCR.
2.LasR PCR.
3.LasR PCR.
-
1.1 kb ladder.
2.Purified P4.
3.Purified LasR.
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Top part is degraded.
Bottom:
1.99 E/P.
2.99 E/P.
3.CI 02 E/P.
4.CI 02 E/P.
09/16/12
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1.-12. psCIC3 omega lysis.
13. 1 kb ladder.
14. psBIc3-GusA.
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Digested A3 and pVeg with E/P.
-
1.ArsR-CzrA 97 E/P.
2.ArsR-CzrA 98 E/P.
3.ArsR-CzrA 99 E/P.
4.02-CI E/P.
5.PCR P4 purified.
09/18/12