Tuesday, May 26, 2015

Fish (Perch) Dissection

Background Information
Where is it found?
The perch is a freshwater fish that is commonly found throughout the British Isles and much of Europe, South Africa, and Asia.
What does it eat?
The perch's diet consists of worms, small fish, and larvae.
How does it breath?
The perch takes in oxygen through its gills which are located on either side of its head. Water enters through its mouth and travels through blood vessels to the gills which filters out carbon dioxide.
Fun fact:
When threatened the perch raises their spiny fins as a defense against being eaten.



Gills- used for breathing 
Swim bladder- contributes to fish's ability to control buoyancy without wasting energy
Liver- acts as a food reserve 
Stomach- digests food
Heart- pumps blood to other parts of body



Use dissecting pins to secure the fish to the dissecting pan. Use scissors to make the cuts through skin and muscle. After making the cuts, carefully lift off the flap of skin and muscle to expose the internal organs in the body cavity.

Grass Frog Dissection

Where is it found?
Grass frogs are found throughout most of North America including Southeastern Virginia, eastern North Carolina, eastern South Carolina, and southern Georgia. The grass frog can be found in meadows, gardens and woodland, and prefer shallow areas of water.
What does it eat?
Frogs tend to prey on insects, worms, snails, and slugs.
How does it breathe?
The grass frog can breathe through its skin.  Because of this, the frog is able to hibernate for months.
Fun fact:
When a male frog successfully attracts a female companion, it will wrap its forelimbs around the female in an embrace.



Liver- secretes bile and processes digested food
Stomach- stores food and begins digestion
Large intestines- stores undigested food
Gallbladder- sac that stores bile made in the liver




Turn the frog on its back and pin down the legs. Cut the hinges of the mouth and open it wide. Use the diagram below to locate and identify the structures inside the mouth. Use a probe to help find each part: the vomerine teeth, the maxillary teeth, theinternal nares, the tongue, the openings to the Eustachian tubes, the esophagus, the pharynx, and the slit-like glottis.

Starfish Dissection


Where is it found?
Starfish are found in saltwater bodies all around the world.
What does it eat?
A starfish's diet usually consists of clams, oysters, sand dollars and mussels.
How does it breathe?
Starfish breathe through their feet since they are made of a thin tissue in which gasses can pass through easily.
Fun fact:
A starfish's mouth is located on the underside of its body while its anus is located on the upper side of its body.



Spines- to provide protection from predators
Ceolomic cavity- holds organs in place
Center dish- center of body containing mouth 





Inside each arm, locate two long digestive glands called the pyloric caeca. These make enzymes to digest food in the stomach. Cut a circular flap of skin from the central disc. (You will have to also cut around the madreporite in order to remove this flap.) Observe the stomach under the central disc. Remove the pyloric caeca from the dissected ray. Find the gonads (testes or ovaries) underneath. These may be small if the starfish is NOT in breeding season. Remove these to see the rest of the water vascular system. Cut off the tip of a ray to observe the parts of the tube feet. Find the zipper-like ridge that extends the length of the ray. The tube feet are attached to these. Locate the bulb-like top of a tube foot called the ampulla. This sac works like the top of an eyedropper to create suction. The bottom of the tube foot is a sucker. Embedded in the soft body wall are skeletal plates called ossicles.

Clam Dissection

'Where is it found?
Clams are found in the coastal saltwater bodies and in shallow freshwater lakes and streams.
What does it eat?
Clams eat plankton.
How does it breathe?
Clam breathe through bivalves that filter their respiration water. Their gills remove oxygen and food particles from the water
Fun fact:
Clams have no eyes, ears, or noses, so they cannot see, hear, or smell.


Gills- used to breathe and move food through body
Foot- can stick out of shell and be used to burrow
Unblock- oldest section of shell
 Mantle- part that forms shell, secretes calcium carbonate 
Cardinal teeth- help shell close properly 


Turn the clam with its dorsal side down and insert a screwdriver between the ventral edges of the valves. Carefully work the tip of the screwdriver between the valves so you do not jab your handTurn the screwdriver so that the valves are about a centimeter apart. Leave the tip of the screwdriver between the valves and place the clam in the pan with the left valve up. Locate the adductor muscles. With your blade pointing toward the dorsal edge, slide your scalpel between the upper valve & the top tissue layer. Cut down through the anterior adductor muscle, cutting as close to the shell as possible.

Grasshopper Dissection

"Where is it found?
It is found in grassy areas worldwide.
What does it eat?
Grasshoppers are herbivores that feed on  a variety of plants.
How does it breathe?
Grasshoppers along with other insects use tracheal breathing.
They exchange oxygen and carbon dioxide between their tissues and the air through a system of tubes.
Fun fact:
Grasshoppers have ears on their abdomen.



Hind leg- 
Abdomen- tail area of a grasshopper (contains heart, digestive system, reproductive organs)
Antennae- sense and touch environment
Hind legs- used to jump 
Walking legs- used to walk 
Thorax- mid area, where legs and wings attach
Wings- used for flying
Eye- made up of hexagonal lenses




Using forceps, remove one of the walking legs and identify these parts --- the coxa connects the femur (the thickest part of the leg) to the grasshopper's body; a slender, spiny tibia connects the femur to the tarsal segments (lowest part of the leg). Raise both pairs of wings and locate the first abdominal segment.

Crawfish Dissection

Where is it found?
Crayfish can be found in any body of freshwater including streams, rivers, lakes, and ponds.
What does it eat?
Crayfish eat fish, shrimp, water plants, worms, insects, snails, plankton, and even dead plants and animals.
How does it breathe?
Crayfish breathe through Internal gills.
Fun fact:
Over 350 species of the 500 crayfish species of the world live in the United States.




Gills- used to breathe under water
Dorsal abdominal artery- central artery coming from heart 
Cardiac stomach- receives food from esophagus for storage
Digestive glands- secrete enzymes for digestion of food



Using one hand to hold the crayfish dorsal side up in the dissecting tray, use scissors to carefully cut through the back of the carapace along the dissection line. Cut along the indentations that separate the thoracic portion of the carapace into three regions. Start the cut at the posterior edges of the carapace, and extend it along both sides in the cephalic region.






Worm Dissection

I'm Where is it found?
Worms are commonly found living in soil.
What does it eat?
Worms get their nutrients from things in the soil such as decaying matter and leaves.
How does it breathe?
Worms must stay moist in order to breathe since they absorb oxygen through mucous on their skin.
Fun fact:
Earthworms are both male and female because they produce both egg and sperm.


Anus- excretes waste
Clitellum- excretes sac that holds eggs
Semenal vesicle- hollow organs that hold semen
Pharynx- muscle that sucks in food
Crop- holds food
Gizzard- breaks down the food
Septa- structural segments
Dorsal blood vessel- main blood vessel towards the rear of worms body 




Position your preserved earthworm dorsal side up and pin it down through the first segment and then again further back behind the clitellum.  Cut a slit in the dorsal surface near the posterior pin.  Using fine scissors extend the cut forward to the first segment.  Be careful not to cut too deep.  Starting at the first segment, cut the septa (thin membranes) that internally divide the segments, so the skin can be laid flat.  Use additional pins to hold the integument open and expose the organs.  Continue to lay the skin back until you have uncovered a centimeter or so of the intestine.

Tuesday, March 17, 2015

Pglo Transformation Lab

Purpose: 
The purpose of this lab is to observe bacterial growth under different conditions and in different environments. 

Introduction:
Genetic transformation is the process of genetic material that is carried by a cell being altered by the incorporation of foreign DNA into its genome. In this experiment, bacteria is going to be transformed by the insertion of a gene that causes the bacteria to glow (pGLO). With this gene, the bacteria should glow green under ultra violet light. Not only does this genetic transformation allow for the bacteria to glow, the pGLO is also resistant to antibiotics. The green fluorescent protein gets switched on by the sugar arabinose. This explains why the bacteria should only glow on plates that contain arabinose on them, but will appear normal on plates without it. 

Methods













Discussion
Gene transformation is when a cell takes in a foreign gene and expresses new DNA. In this lab, we infused the Green Fluorescent Protein (GFP) from a jelly fish along with an antibiotic resistant gene into the DNA of E. coli with the help of a plasmid. Plasmids are circular DNA that usually contain genes for one or more traits. We used the pGLO plasmid which contained the gene for GFP and a gene for resistance to the antibiotic ampicillin as mentioned above. In this lab we had 4 dishes. The first was our control group which held bacteria only provided with LB (food). This plate had growth because it was given food, however the bacteria didn't glow since it was not exposed to the pglo gene. The second plate was al so a control group. It had LB and ampicillin which is an antibiotic (kills bacteria). Therefore there was no growth on this plate because the genes were not exposed to the antibiotic resistant gene. The third plate had LB, and ampicillin, however it was exposed to the plasmid which contained the antibiotic resistant gene. So there was growth, but less than the LB control plate because only some of the bacteria took in the antibiotic gene into their DNA. The last plate had LB, ampicillin, and the pglo gene so there was growth because it had the antibiotic resistant gene, and the bacteria glowed in the dark. Since it took up the pGlo gene.

Conclusion
Overall, we learned how we can mash two different types of DNA together. It was cool to see that just through a simple experiment we could make a bacteria become antibiotic resistant and glow in the dark. Gene transformation is being used recently by scientists to experiment putting specific genes in bacteria to treat certain diseases. It was cool to think that we could do an experiment that professionals are using now to advance science and medicine. 











Monday, March 9, 2015

Restriction Mapping of Plasmid DNA

Purpose:
The purpose of this lab was to experiment with the usage of restriction enzymes on plasmid DNA using gel electrophoresis.

Introduction:
A restriction map is a visual of DNA with known restriction sites of that given sequence. A restriction map requires the use of restriction enzymes, which are enzymes that chop up a DNA molecule at a specific site. These DNA fragments are then separated by argos gel electrophoresis.  The distance between restriction enzymes can be found by comparing the patterns of the fragments. This process can be used to figure out information about the structure of an unknown piece of DNA. The fragments are compared to a lane of DNA markers which works a a size comparison chart. In this way the size of a DNA segment (number of base pairs) can be estimated.

Procedure: 

1) Carefully insert the buffer unto the gel's well by squeezing the pipette. Use the following table to make sure each buffer is in its correct well.
2) Bring the gel, which should look like the one below, to the electrophoresis area made by your teacher and place into the solution. 


3) The next day, take your gel out of the solution in the electrophoresis chamber and place it on a light box. It may help to mark the visible bands with some kind of marker to make them easier to see.







Discussion: 
PstI ad bands at 700 and 4700. PstI/ SstI had bands at 700, 2200, and 2500. PstI/ HpaI had bands at 700, 500, and 4200. PstI/ SstI/ HpaI had bands at 700, 1300, and 3000. Each digest added up to 5400 and had a band at 700. Because HpaI only has two fragments it must be circular. If the DNA were linear, there would be three different fragments with two different cites. There is only one PstI site because we know the HpaI has two cites already and there are only three fragments of DNA in the sample PstI/ HpaI. With circular DNA each site produces one fragment. We know SstI only has one site for the same reasoning mentioned before. Based on the position of the fragments on the gel, the 700 base pair fragment of HpaI is  unaffected by the two other enzymes. There is no fragment that appears only in the HpaI/PstI/SspI digest. This means that the  enzymes of HpaI, PstI, and SspI spliced the DNA in their respective locations/codes. Because  there are no other fragments present, HpaI, PstI, and SspI were the only restriction enzymes  used to digest this enzyme. 

Conclusion:
The purpose if this lab was to find out the number of cut sites present in the DNA sequence for each restriction enzyme and the position of the cuts relative to one another. We were successful in achieving a satisfactory result through the use careful observations and calculations. We were able to font the number and locations of all of the cut sites.

Saturday, February 14, 2015

How to Extract DNA from a Strawberry

How does it work?
Strawberries have more DNA than any other fruit with 8 pairs in each chromosome. To extract the DNA, each of the solutions used in the experiment plays a part. The soap dissolves the cell membrane, which frees the chromosomes from the nuclear membrane. The salt is added to break up the nucleotides in the DNA sequence. Finally, DNA is not soluble in alcohol because it is polar, and even less so when the alcohol is chilled.

Materials-
A strawberry
A plastic bag
10 ml of soapy salt water
100% alcohol solution
Pipett
A coffee filter
A clear test tube

Procedure
1) Rip off the green part of the strawberry
2) Put strawberry in plastic bag and mush up until pulped
3) Add 10 ml of the soapy solution to the bag, seal, and then continue to squish some more. 
4) Put cone into test tube, and put coffee filter into the cone
5) Slowly pour strawberry gunk into coffee filter
6) Allow the strawberry juice to filter through the coffee filter, it will take a few minutes
7) Add 2 ml of cold 100% alcohol to the strawberry juice
8) Watch for a minute and the line between the red juice and the clear solution will become foggy.
9) Put stirrir into foggy area and twirl
10) Pull out the stirrer, see that snot looking stuff? That's DNA!