Illustration of sexduction in E. coli using streptomycin resistant F-lac- recipients and F′lac+ donors
Abstract
The process in which DNA from a donor cell is transferred through physical contact into a recipient cell is known as conjugation. In the experiment, sexduction, a cross between F′ and F- plasmids, was demonstrated by mixing the donor and recipient, diluting them and using selective plating methods. The plasmids from the donor, recipient and transconjugant were isolated and analyzed using FIGE gel Electrophoresis. The data that was acquired consisted of 12 red colonies and 483 white colonies from the conjugation mixture with a conjugation efficiency of 2.42%. The FIGE gel Electrophoresis produced the DNA bands of the donor and transconjugant strains with a size of 112 kb and no bands for the recipient. The results show that the recipient cells were converted to donor cells by the transfer of the F′ plasmid therefore, sexduction must have occurred.
Introduction In 1946, conjugation was first discovered by Lederberg and …show more content…
The supernatant was discarded and 1mL of 70% ethanol was added and centrifuged for a minute and removed. Once the pellet was completely dry, 20 µL of TE was added and mixed. Once fully mixed, 4 µL of the gel loading dye was added. The BIO-RAD FIGE Mapper apparatus was prepared by the instructor. The entire sample of Plasmid DNA was loaded into a well of the gel. The instructor conducted the apparatus until the gel program was finished. The Field Inversion Gel Electrophoresis in 1% pulse field certified agarose applying 180 V forward and 120 V reverse, with the help of Bio-Rad FIGE Mapper using program 4 for 16 hours. The gel is placed in an ethidium bromide solution for 15 minutes and transferred to distilled water for 30 minutes. It is examined under UV light box and then a picture of the gel was taken using the Kodak EDAS
Abstract
The process in which DNA from a donor cell is transferred through physical contact into a recipient cell is known as conjugation. In the experiment, sexduction, a cross between F′ and F- plasmids, was demonstrated by mixing the donor and recipient, diluting them and using selective plating methods. The plasmids from the donor, recipient and transconjugant were isolated and analyzed using FIGE gel Electrophoresis. The data that was acquired consisted of 12 red colonies and 483 white colonies from the conjugation mixture with a conjugation efficiency of 2.42%. The FIGE gel Electrophoresis produced the DNA bands of the donor and transconjugant strains with a size of 112 kb and no bands for the recipient. The results show that the recipient cells were converted to donor cells by the transfer of the F′ plasmid therefore, sexduction must have occurred.
Introduction In 1946, conjugation was first discovered by Lederberg and …show more content…
The supernatant was discarded and 1mL of 70% ethanol was added and centrifuged for a minute and removed. Once the pellet was completely dry, 20 µL of TE was added and mixed. Once fully mixed, 4 µL of the gel loading dye was added. The BIO-RAD FIGE Mapper apparatus was prepared by the instructor. The entire sample of Plasmid DNA was loaded into a well of the gel. The instructor conducted the apparatus until the gel program was finished. The Field Inversion Gel Electrophoresis in 1% pulse field certified agarose applying 180 V forward and 120 V reverse, with the help of Bio-Rad FIGE Mapper using program 4 for 16 hours. The gel is placed in an ethidium bromide solution for 15 minutes and transferred to distilled water for 30 minutes. It is examined under UV light box and then a picture of the gel was taken using the Kodak EDAS