pGLO Transformation Lab

Hello and welcome to our fabulous P.Glo lab done by Aliya, Emma, Cara, and Ally (your fav senior ladies who value and appreciate you Mr.Filipek) 

Purpose

The purpose of this experiment was to perform and prove genetic transformation which is insertion of a gene into an organism in order to change it's trait. In this experiment we were testing the insertion of the GFP(green fluorescent protein) gene into the P.Glo plasmid of the E.coli cells. 

Introduction

In this experiment, E.coli is a bacteria that is commonly found inside of mammals and birds and is used to breakdown food into small molecules that can be transferred into your blood. It is a very basic prokaryotic cell and only lives a short time. Most types of E.coli are good for you but there are a very few select types that can be poisonous to humans such as E.coli found in raw meat. Genetic transformation, in this experiment is used to transfer new DNA from the GFP to plasmid of the E.coli cells. A plasmid is a small circular piece of DNA where the GFP can be inserted in order to show new traits within the cell. The insertion of GFP into the pGlo makes the cell resistant to antibiotics.

Methods

We started of by using two micro test tubes one of +pGlo (containing pGlo) and one of -pGlo (not containing pGlo). We then placed 250 microliters of CaCl2 into each tube. Next the two tubes we're placed on ice. After this, we put a sample of the E.coli bacteria into each tube using a loop. Immediately, we inserted pGlo plasmid DNA into the positive pGlo tube but not in the negative pGlo tube. Then we incubated (let them sit on ice) for 10 minutes. Right after this, we performed a heat shock on both tubes. Next, we put 250 microliters of LB nutrient broth in both tubes. We then placed 100 microliters of the solution from each tubes onto 2 Agar plates each, resultin in 4 total plates. One plate of +PGlo LB/amp, one of +pGlo LB/amp/ara, one of -pGlo LB/amp, and one of -pGlo LB.

Data

Graphs and charts

Drawn-out representations of our Agar plates and their contents after the lab was completed

Discussion

Our results from this lab were a direct result of the proccess of genetic transformation. Some of the dishes expressed the pGLO trait while others did not because of this proccess. Those that did express the trait those that expressed the "glow in the dark" trait were those with pGLO and LB/amp. The plate with LB/amp/ara contained more  colonies because of the sugar that was added to the plate that helped with growth. If we were to make adjustments to this lab, we would put more of each substance of each plate in order to see more colonies then what we did the first time. It's difficult to tell whether our results are completely accurate due to how little of data that we have. As predicted, the petri dishes that had the GFP inserted into the plasmid glowed in the dark, while the petri dishes without the plasmid did not. The insertion of the GFP into the plasmid then makes it resistant to antibiotic. The LB plate has no growth because it has no protein to resist antibiotic. 

Conclusion 

Through this lab we were able to prove that pGLO causes the bacteria to be able to resist the antibiotic. 

References 

http://scienceforkids.kidipede.com/biology/cells/ecoli.htm

Pre-lab selfie (super excited)

Receiving the translation solution (super fun)

Icing the solutions (super chill) 

Getting the E.coli (super thrilling)

E.COLI (yay)

Heat killing the solutions (super amazing)

Aliya and Emma in the action (feat. Tape and coffee)

Petri dish! (Super scientific)

Mixing! (Super cool and fun)

All the Petri dishes in their final stage of the lab! ( super exciting and biological) 

+pGlo LB/amp/ara

-pGlo LB

+pGlo LB/amp

-pGlo LB/amp

How To: Extract DNA From a Strawberry

Purpose: Hello! This was one of our favorite labs because we all love strawberries a whole lot. The purpose of this experiment was to see if there was a way to extract a strand of DNA from one whole fresh strawberry. 

Introduction: DNA is found in all living organisms and in this lab our goal was to extract a strand/fibers of DNA from the strawberry. 

Methods: First we mashed up the strawberry in a ziplock bag in order to make the DNA more extractable. Then we mixed the mashed up strawberries with the extraction buffer (the soapy water mix). After letting the mixture soak through a coffee filter into a graduated cylinder, we collected the liquid after a few minutes. We then added a few drops of ice cold alcohol to the strawberry and soapy water solution. We made sure to let the alcohol drip down the side of the graduated cylinder instead of dropping it straight in. After a minute or two a white cloudy layer appeared. This was the DNA! After stirring the pipet in the mixture, the DNA climbers could be pulled out and examined.  

Discussion: Our results from this lab successfully proved this lab true. DNA could be successfully extracted from a strawberry as long as we followed the correct steps, which we eventually did. The only thing that went wrong was our first attempt at the lab. We first mixed the mashed up strawberry with the ice cold alcohol to find nothing so we knew then that we had to approach the lab differently. Eventually we figured it out and successfully extracted the DNA from the strawberry. If we were to improve our tactics and techniques in this experiment, we would have not first mixed the alcohol with the strawberry solution. The results of our experiment proved that DNA fibers/strands are found in strawberries. 

Conclusion: To conclude this experiment, DNA can be extracted from raw strawberries as long as it is mixed with an extraction buffer, filtered through a coffee filter and mixed with alcohol. Our results from our lab support this conclusion. 

References: 

DNA Extraction From Straberries (Schoology) 

Pictures

The mashed up strawberries!!!

Adding the alcohol to the solution!!!

Extracting the DNA (feat. Mr. Filipek) !!!