Essay about Biology Lab

Step 4: Place the test tubes into separate coffee cups to maintain the upward position. Add 2 mls of the catalase solution to each of the test tubes and then place tubes1, 2, and 3 in the conditions described above. For test tube 4, fill the coffee mug half full of boiling

2. Once your solutions are made and set aside, you will want to create your simulated cells using dialysis tubing. Fold one end of the dialysis tubing and wrap a rubber band (tightly) around that end to seal it.

items presently in solid form in the lab kit or purchased separately for this laboratory

Cells are rinsed with chilled PBS, scraped into chilled PBS, and then centrifuged at 200 × g for 4 min. Discard the supernatant. The cells are now ready for lysis or storing at -80 ℃ for future use.

This lab is about moving genes from one thing to another using plasmids. Plasmid has the ability to replicate, so it replicates independently, and separately from the chromosomal DNA. Plasmid are one or more small piece of DNA and they enter cells as a double strand DNA. When they enter the cell as a doubke strand they do not invade he chromosomal DNA. We will also transform bacteria into GFP which is mainly from the jelly fish Aequorea Victoria. The GFP causes the the jelly fish to fluorescent and glow in the dark. After the transformation, bacteria starts to make the GFP which causes them to glow a green color under a ultraviolet light.

The pGLO plasmid is engineered to express green fluorescent protein (GFP) in the presence of the sugar arabinose as well as the ampicillin resistance gene β-lactamase (bla) (Brown, 2011). Original E. coli HB101 do not have ampicillin resistance or the GFP gene allowing them to glow under UV light. In this experiment, we transformed E. coli HB101 with the pGLO plasmid by heat shock to make the bacterial cells competent, allowing the plasmid to enter the cell (Brown, 2011). The mixture of bacteria with pGLO plasmid were given recovery time after heat shock, then spread on LB/amp and LB/amp/ara agar plates. The bacteria mixture with no plasmid added were spread on LB and LB/amp agar plates and all four plates were incubated at 37°C for

room temperature. Vials 4, 5, and 6 should be in the bath containing water that is 10oC. After the

What is the goal of this lab? What question is it trying to answer, or what problem is it trying to explain?

Next heat shocks the tubes for 50 seconds, followed by icing for 10 more minutes. The heat shock increases the permeability of the cell membrane to DNA. Then add 250 milliliters of LB and incubate for 20 minutes. The 20 minute incubation following the addition of LB broth allows the cells to grow and express the ampicillin resistance protein, beta-lactamase, so that the transformed cells survive the subsequent ampicillin selection plates. Plate 100 microliters the + tubes evenly on two plates; 1 of LB and Amp, and one of LB, AMP, and ARA. Plate 100 microliters of the – tubes evenly on two plates; 1 of LB and AMP, and one on LB only.

The purpose of the experiment was to isolate plasmid DNA, followed by restriction digestion using restriction endonucleases and then visualizing the digested fragments after subjecting to gel electrophoresis. Plasmid DNA (pSP72 DNA) was isolated from Escherichia coli KAM32 (E.coli) cultures using the QIA prep miniprep kit and then subjected to restriction digestion by EcoRI and HindIII. The restriction digested DNA was then loaded into the wells of 0.7% agarose gel and subjected to electrophoresis. It can be concluded from our results that our plasmid DNA isolation was successful and the restriction digestion results were partially in agreement with our hypothesis.

Next, gather 0.5 ml of the protein extract and save this portion in a separate container. This will be used for the preparation of the membrane.

Before plating the strains on agar plates, dilutions of the three strains of cells were prepared with LB broth.

The overall goal of the DNA lab was to create recombinant plasmids which we would use to alter the DNA of bacteria to make them ampicillin resistant. The first test of our success was the gel electrolysis. The gel photo allowed us to see if we had successfully ligated the recombinant plasmids. For the uncut samples, K- and A-, we expected to see two or three bands, since there are several forms of plasmids: supercoiled, nicked or open-circle, and the multimer. In K- there were two bands, and in A- we saw three bands, indicating that all three forms of plasmids were present.

3. Use a sterile pipette to transfer 0.1 ml of each dilution on to a MacConkey agar plate.

7.When water bath is ready, put each test tube into the water bath. Wait 5 minutes.