User:Eduardo Vladimir Munoz/Notebook/UNAM Genomics Mexico 2011/2011/09/24

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High Stabilitiy Plasmid Development

Abstract

  • Plasmid extraction of the B0015 dephosphated and repC ligation
  • Gels to corroborate the identity of strains 2, 10 and 11


Extractions

Two strains (labeled 3 and 5) were inoculated into liquid LB media and grown for plasmid extraction.


Electrophoresis gels

The digestions gels had not enough concentration to corroborate the positives. Yet, the PCR amplification revealed that the repC region is only contained in strains 2 and 10.

ElectrophoresisGel24Sep2011PCRControlRepCter.JPG
1 Ladder 500bp
2 C+ PCR
3 C-
4 strain 2 True Positive
5 strain 10 True Positive
6 strain 11 False Positive


ElectrophoresisGel24Sep2011Digestions EcoRV XhoI.JPG
1 Ladder 500pb
2 B0015 in pSB1AK3
3 B0015 in pSB1AK3 EcoRV
4 B0015 in pSB1AK3 XhoI
5 repCter strain 2
6 repCter strain 2 EcoRV
7 repCter strain 2 XhoI
8 repCter strain 10
9 repCter strain 10 EcoRV
10 repCter strain 10 XhoI
11 repCter strain 11
12 repCter strain 11 EcoRV
13 repCter strain 11 XhoI
14
15


ElectrophoresisGel24sep2011 repCterDigestionsRepetition.JPG
1 Aarón sample
2 Fermentas ladder
3 B0015 in pSB1AK3
4 B0015 in pSB1AK3 EcoRV
5 B0015 in pSB1AK3 XhoI
6 repCter strain 2
7 repCter strain 2 EcoRV
8 repCter strain 2 XhoI
9 repCter strain 10
10 repCter strain 10 EcoRV
11 repCter strain 10 XhoI
12 repCter strain 11
13 repCter strain 11 EcoRV
14 repCter strain 11 XhoI
15