Boston UniversityCompTransf
From 2007.igem.org
(→Making Shewanella Competent and Transforming Plasmid) |
|||
| Line 5: | Line 5: | ||
Zymo is a company that sells a package of wash buffer, competent buffer and dilution buffer. Using the protocol listed on this website, shewanella was made competent, meaning able to take in a plasmid. | Zymo is a company that sells a package of wash buffer, competent buffer and dilution buffer. Using the protocol listed on this website, shewanella was made competent, meaning able to take in a plasmid. | ||
| - | The plasmid pJQ200 was transformed into shewanella | + | The plasmid pJQ200 was transformed into shewanella. Since this plasmid has a gentamycin resistance gene, the shewanella was plated on gentamycin plates. After overnight incubation the plates were filled with bacteria, proving that the bacteria had taken in the plasmid and gained resistance due to it. |
To confirm that the plasmid was definitely pJQ200, a portion of bacteria was plated on | To confirm that the plasmid was definitely pJQ200, a portion of bacteria was plated on | ||
a 10% sucrose and gentamycin plate. No cells grew after incubation, as expected if transformation worked correctly due to pJQ200's sacB gene, which acts as a suicide vector in the presence of 10% sucrose, killing the bacteria housing it. | a 10% sucrose and gentamycin plate. No cells grew after incubation, as expected if transformation worked correctly due to pJQ200's sacB gene, which acts as a suicide vector in the presence of 10% sucrose, killing the bacteria housing it. | ||
Revision as of 03:17, 4 July 2007
Making Shewanella Competent and Transforming Plasmid
Zymo is a company that sells a package of wash buffer, competent buffer and dilution buffer. Using the protocol listed on this website, shewanella was made competent, meaning able to take in a plasmid.
The plasmid pJQ200 was transformed into shewanella. Since this plasmid has a gentamycin resistance gene, the shewanella was plated on gentamycin plates. After overnight incubation the plates were filled with bacteria, proving that the bacteria had taken in the plasmid and gained resistance due to it. To confirm that the plasmid was definitely pJQ200, a portion of bacteria was plated on a 10% sucrose and gentamycin plate. No cells grew after incubation, as expected if transformation worked correctly due to pJQ200's sacB gene, which acts as a suicide vector in the presence of 10% sucrose, killing the bacteria housing it.
