BSC1010L SectionU42
The effect of Control Plasmid and Plasmid Lux in Bacterial Transformation
Vanessa Louis
PID: 4709178
Lab Partners:
Patrick Abaunza
Raquel Alespeiti
Robert Lugo
Date Performed:
31 October 2014
TA: Reinier Llorca
Abstract
According to The General Biology I Lab manual, involved in Gene expression are two complex cellular pathways, each being subjected to regulation (Alberte et al., 2012). The first pathway is the synthesis of an RNA molecule from a DNA strand. This process is called transcription. When the RNA molecule exits the nucleus, it can be translated into a polypeptide sequence. This process is called translation. With some adjustments, these pathways happen in both prokaryotes and eukaryotes. Plasmids …show more content…
We were given three agar plates by our TA, each agar plate containing a nutrient broth. We labeled each agar plate with whatever content we were going to put inside of it. We worked with the control so our three labels were “LB^NP, LB^AmpC, and LB^C”. LB^NP means the broth with no plasmid. LB^AmpC means the broth with ampicillin and the control plasmid. Last but not least, LB^C means the broth with just the control plasmid. We prepped our cell spreader for its use to come before we needed it. Patrick and Robert lit the ethanol lamp and let it burn for a couple seconds. They dipped the cell spreader in ethanol and passed it cell spreader across the flame over the ethanol lamp. Raquel and I continued to work with the bacteria and agar plates. I used a sterile pipet and removed 130 µL of that mixed solution from the control tube and dispensed the bacteria onto the agar plate that labeled LB^AmpC. Soon after I dispensed it, Patrick used the cell spreader to spread the bacteria evenly onto the agar surface. We repeated the same process for the agar plate that labeled LB^C. For our last plate, we pipetted 130 µL from the tube that labeled no plasmid and put it in the plate that was labeled LB^NP. Patrick came along soon after and used the cell spreader one last time to spread the bacteria evenly among the …show more content…
The two types of plasmids are a control plasmid and Lux plasmid. I also tested for luminescence in the bacteria after being incubated for 7 days. I hypothesized that if there was plasmid added to the bacteria in the agar plate, that it would have colonial growth or no growth at all. If there was no plasmid added, with no specifications to what type of plasmid, the bacteria would have lawn growth with no restrictions at all. I came to this conclusion for my prediction because according to one of my sources which is an article in the ISME journal, plasmids serve as main vessel that allows gene flow, linking different genetic pools. I translated that to be that plasmid would help it grow but ampicillin would still restrict
The effect of Control Plasmid and Plasmid Lux in Bacterial Transformation
Vanessa Louis
PID: 4709178
Lab Partners:
Patrick Abaunza
Raquel Alespeiti
Robert Lugo
Date Performed:
31 October 2014
TA: Reinier Llorca
Abstract
According to The General Biology I Lab manual, involved in Gene expression are two complex cellular pathways, each being subjected to regulation (Alberte et al., 2012). The first pathway is the synthesis of an RNA molecule from a DNA strand. This process is called transcription. When the RNA molecule exits the nucleus, it can be translated into a polypeptide sequence. This process is called translation. With some adjustments, these pathways happen in both prokaryotes and eukaryotes. Plasmids …show more content…
We were given three agar plates by our TA, each agar plate containing a nutrient broth. We labeled each agar plate with whatever content we were going to put inside of it. We worked with the control so our three labels were “LB^NP, LB^AmpC, and LB^C”. LB^NP means the broth with no plasmid. LB^AmpC means the broth with ampicillin and the control plasmid. Last but not least, LB^C means the broth with just the control plasmid. We prepped our cell spreader for its use to come before we needed it. Patrick and Robert lit the ethanol lamp and let it burn for a couple seconds. They dipped the cell spreader in ethanol and passed it cell spreader across the flame over the ethanol lamp. Raquel and I continued to work with the bacteria and agar plates. I used a sterile pipet and removed 130 µL of that mixed solution from the control tube and dispensed the bacteria onto the agar plate that labeled LB^AmpC. Soon after I dispensed it, Patrick used the cell spreader to spread the bacteria evenly onto the agar surface. We repeated the same process for the agar plate that labeled LB^C. For our last plate, we pipetted 130 µL from the tube that labeled no plasmid and put it in the plate that was labeled LB^NP. Patrick came along soon after and used the cell spreader one last time to spread the bacteria evenly among the …show more content…
The two types of plasmids are a control plasmid and Lux plasmid. I also tested for luminescence in the bacteria after being incubated for 7 days. I hypothesized that if there was plasmid added to the bacteria in the agar plate, that it would have colonial growth or no growth at all. If there was no plasmid added, with no specifications to what type of plasmid, the bacteria would have lawn growth with no restrictions at all. I came to this conclusion for my prediction because according to one of my sources which is an article in the ISME journal, plasmids serve as main vessel that allows gene flow, linking different genetic pools. I translated that to be that plasmid would help it grow but ampicillin would still restrict