Transformasi Fragmen DNA Salmonella Ke Dalam Sel E. coli DH5-α
| Main Authors: | Rahayu, E.S., Pertiwiningrum, A., Indrati, R., Raharjo, S., Rahayu, S., Margino, S. |
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| Format: | Article info application/pdf |
| Bahasa: | eng |
| Terbitan: |
Faculty of Agricultural Technology, Universitas Gadjah Mada, Yogyakarta, Indonesia
, 2017
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| Online Access: |
https://jurnal.ugm.ac.id/agritech/article/view/19320 https://jurnal.ugm.ac.id/agritech/article/view/19320/12546 |
Daftar Isi:
- Genomic DNA from Salmonella typhimarium ATCC 14028, cultured in a Luria broth for 24 hr at 37°C, was isolated from the cells using EDTA as a chelating agent and SDS as a detergent and lysing agent. The amount of genomic DNA was then digested with each of three restriction endonucleases, i.e., EcoRI, HindlII, and BamHI for 4 hr at 37°C. A plasmid (pUC19) was used as a vector. Prior to use, the pUC19 plasmid was split with one of the three restriction enzymes and dephosphorylated using a bacterial alkaline phosphatase. The genomic DNA was then ligated to the corresponding prediggested plasmid using a ligase at 15°C over night. Transformation of the DNA recombinant into E.coli DH5-a was successfully carried out using a heat shock method as indicated by gel electrophoresis