HOW TO: Dissect a Starfish

Starfish (Sea Star)

Background

These invertebrates reside in all parts of the ocean, including the Atlantic Ocean to the Indian Ocean. Starfish primarily feed on clams, oysters, sand dollars, mollusks and mussels. Starfish are able to breathe through their feet. Their feet are made of a very thin tissue in which gases can move easily. Ironically, starfish are not actually fish. They do not have gills, fins, or scales like fish do. They move with their feet while fish propel themselves through the water.

External Anatomy

Aboral Surface— side of the starfish without a mouth ("top")

Oral Surface— side of starfish with mouth ("bottom")

Eyespot— lies underneath the skin, can detect light and dark

Central Disc— center of the starfish from which the arms start from, contains the madreporite, mouth, and anus

Madreporite— used to filter water into the water vascular system of the echinoderms

Ambulacral Groove— contains the feet on the oral side and used to open up shells for food

Internal Anatomy 

Digestive glands— break down food by the use of enzymes and connected to the stomach

Ring Cartal— carries water from the stone canal to the radial canal, connects the two canals

Ossicles— forms part of the exoskeleton and provides protection and strength

Gonads— reproductive organs (ovaries or testes) that are used with reproduction by producing egg or sperm

Lateral (Radial) Canal— runs the length of the starfish arm and is part of the water vascular system

Ampulla— fills up with water and then releases water to feet

Incision Guide

Dissection Procedure

 

HOW TO: Dissect a Grasshopper

Grasshopper (Romalea)

Background

Grasshoppers are widespread in the United States, they are most prominent in the fall months. Found in areas with grasses, vacant lots, and gardens. Grasshoppers are herbivores so they manly feed on grasses. Grasshoppers have holes on their abdomens and thorax called spiracles. They have an exchange of oxygen and carbon dioxide between their tissues (called tracheal breathing). In the early stages of a grasshopper's life, they do not have wings. However, as they mature, they develop wings. They are visible as small pads at the end of the thorax. 

External Anatomy

Head

Antennae - located on the head above the mouth.  Is used like a human's nose, but senses odors, touch, humidity, vibration, wind velocity, and wind direction.

Compound eyes - (2) can see shape, color, movement and distance.

Simple eyes - can only detect light intensity

Thorax

Legs - There are both walking and leaping legs.  The four walking legs are located toward the front of the thorax, and the two leaping legs are located toward the back.

Abdomen 

Ovipositer (females) - Where tube is located to deliver eggs

Spiracles - used to breathe

Internal Anatomy 

Dorsal Blood Vessel - heart

Tracheae - network of air tubes

Ganglia - bundle of nerve cells

Crop - stores food

Gizzard - grinds food with chittinous plates

Intestine - transports waste through the digestive system from the ind gut to the rectum

Malpighian Tubules - removes chemical waste from the blood

Rectum - passageway for digested waste from the intestine to the anus

Incision Guide

Dissection Procedure

HOW TO: Dissect a Clam

Clam (Bivalvia or Pelecypoda)

Background

Tend to inhabit shallow waters, there is over 15,000 different living species and 500 live in fresh water while the rest live in salt water. Clams are filter feeders, they pump water through their bodies to capture microscopic organisms such as plankton. The clam breaths through gills, the gills are used for both breathing and feeding the clam. The clam sticks the ends of its siphons out of the shell and food and oxygen come in one end and out the other. Clams move by the use of their foot, the foot allows the clam to move from side to side when necessary. Below is a video of a clam using its foot in order to move on land:

External Anatomy

Umbo— oldest part of the clam, where the clam, highest part of the shell

Growth Lines— shows the age of the clam

Hinge Ligament— hinges the valves together and allows them to open and close, important part

Internal Anatomy

Stomach— separates water from the food, starts digestive process 

Gill— filters oxygen from the water in order for the clam to breathe

Mantle— cover for the clam that helps with growth, respiration, and shell color

Axe foot— allows the clam to burrow and move around, axe shaped

Incision Guide

Dissection Procedure

Restriction mapping of plasmid DNA

Purpose: The purpose of this lab was to characterize a DNA sequence.

Introduction: A plasmid is a small DNA molecule that's separated from a chromosomal DNA and can replicate on its own.  Restriction mapping is obtaining the structural information of DNA by using restriction enzymes. These restriction enzymes cut DNA at specified regions called "sites". 

Methods: We started out by adding the dye into each designated slot of our gel. We were to aim the pipet into the groove without puncturing the gel. If we punctured the gel or placed the gel in the opposite way, this would have ruined our experiment. After adding the dye to each slot, we let it sit for a day so the restriction enzymes could cut at their specified region. The next day, we came back and measured how far each DNA region traveled down the gel. 

Data

Graphs and charts

Discussion 

    Our results from this lab came out to be very close to what they should have been.  The only aspect of our results that did not come out ideally is the number of cuts that were made in the final two columns.  While there should have been three distinct cuts in the fifth column, and four in the sixth.  Our gels only showed two cuts in the fifth column and three in the sixth.  A potential reason for this outcome may be that we simply could not see the cuts due to a lack of pigment in the gel.  This pigmentation issue was probably derived from the beginning of the experiment when the liquids were being placed in the gels.  If any amount of the contents from the reaction tubes escaped during the piper ting process, it would make the fragments more difficult to detect.  This means that when loading the last two wells, a large amount of the liquid escaped, and therefore we could not see the extra fragments.  In the future, we would be more careful and precise when transferring the fluid from the reaction tubes to the wells. When labeling our fragments based on size, we knew that the larger fragments didn't travel very far, versus the smaller fragments, which went the largest distance.  With this in mind, we made general estimations on our own gel.  In doing this, it is possible that the lengths we assigned to our fragments may be off.  However, when considering the data, it is all compared to each other, so there should not be much wrong with what information we came out with in the end.  All in all, this was a successful experiment.

Conclusion

To conclude our lab, restriction map information is important for many processes used to manipulate DNA. In this lab for example, one application was to cut a large strand of DNA into smaller pieces to allow it to be sequenced. We were successfully able to characterize a DNA sequence through this process. 

Gel map artfully drawn out by Mr.Filipek (super artistic and well-done)

Emma adding the DNA loading dye to our gel (super successful)

Our whole classes gels all together in the electrophoresis chamber (most of them look amazing good job class)

Our lab group's gel after the electrophoresis was complete 

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) !!!

Cell Communication Lab

Purpose

The purpose of this experiment was to observe cell communication between two strains of yeast (A-type and Alpha-type), both of the two being opposite sexes.

Introduction

Cell Communication is essential for organisms in order for signals to turn into responses within the cell. In this lab we will be studying methods by which yeast cells partake in cell communication. It is our job to determine what type of cell signaling  these yeast cells use and how these cells send signals to each other. We will complete this lab with the use of yeast, Petri dishes, and microscopes. 

Methods

We began this lab by separating samples of the a-type, the alpha-type, and a mixture of the two in order to determine the reproduction rate and how the yeasts go about it asexually as well as sexually. We placed each of the wet yeasts in its own Petri dishes and let them sit over night. Before we wrapped up the lab on the first day we measured the yeasts under a microscope. We put broth in the test tubes and placed them into an incubator overnight,. The next day we scraped the dried yeast off the petri dishes and mixed if with water and put each yeast on a glass slide and put it under a microscope. We then measured our data to find the change in yeast cells over time. 

Mixed Yeast (Day 1)

A Type Yeast (Day 1)

Alpha Type Yeast (Day 1)

Alpha-Type Dry (Day 2)

Mixed Dry (Day 2)

A-type (Day 2)

Mixed Wet (Day 2)

Data

Mixed Yeast Culture Data

Alpha-Type Yeast Culture Data

A-Type Yeast Culture Data Table

Graphs and Charts

Graph of the percent change of yeast cells over time (haploid and diploid)

Discussion

In the Cell Communication Lab we measured the difference between the different yeasts (alpha type, A-type, and mixed) to see their sexual/asexual production. We observed that there was not much difference between the a-type and the alpha-type strains of yeast because they are both able to reproduce asexually, there was not much difference in budding of the cells. But, when the alpha-type and A-type were mixed together it is observed that right away there is already diploid zygotes and diploids zygote budding from the cells beginning to grow towards each other because they are opposite sexes. This goes to prove that though yeasts are able to reproduce asexually, bringing the two opposite sexes together causes cellular communication to occur by the sending and receiving of signals to reproduce at a more rapid rate. One thing that could be done differently in this experiment was counting of the different haploid and budding haploid cells. More test trials would have been more efficient but we spent a majority of time trying to get pictures to accurately represent our data. Another thing would be to use different settings on the microscope to get a closer look at the cells and be able to examine the mixture culture closer. 

Conclusion

In this experiment we see that the most favorable condition for yeast reproduction is when yeasts are mixed with a-type strains and alpha-type strains. This is because of the cell's ability to to communicate through signaling between different sexes and as long as the environment is favorable, diploid zygotes are formed and divide.

References