Dissection

http://youtu.be/PufiqmxGiYU

http://youtu.be/XglaL4QxEr0

Gel Electrophoresis Lab

 

In this lab, we took a sample of DNA and performed gel electrophoresis.  Gel electrophoresis is used to create restriction maps and identify DNA.  Here's what we did:

Methods:

We took 5 samples of DNA and used three separate restriction enzymes to cut it.  Each sample of DNA contained a combination of restriction enzymes, and then was placed into separate wells on the electrophoresis gel.  This is what each sample of DNA contained:

1. Marker (Only contained DNA)
2. DNA + PstI
3. DNA + PstI/HpaI
4. DNA + PstI/SspI
5. DNA + PstI/HpaI/SspI

Once each sample was placed into the gel wells, the entire gel box was placed into the gel electrophoresis machine.  Electricity is run through the gel, propelling the light (smaller) strands a DNA far away from it's original position, and propelling heavy (bigger) strands of DNA a smaller distance from it's original position.  We then tallied the size of each strand of DNA based on it's distance from the origin, and also by comparing the DNA strands to the marker in well 1.

Discussion:

By observing the positions of the strands in each well in relation to the marker, we were able to approximate the length of each strand.  When all of the strand lengths added up to 2,600 base pairs in well two, we then knew that wells three, four, and five must also have the same number of base pairs.  We were able to create restriction maps containing each restriction enzyme according to the length of DNA strands and number of base pairs in each strand.  For example, well two contained two fragments of 900 and 4700 base pairs. so therefore there must have been two restriction sites, creating two sections of DNA made of 900 and 4700 base pairs.  We were able to perform the same action on well three and four, creating restriction maps for each of their restriction enzymes, respectively.  In order to make a restriction map of DNA cut by all three enzymes, we simply superimposed each map from well two, three, and four on top of eachother.    

Here is an image of the number of base pairs per DNA strand

                                    

Here are our restriction maps for each enzyme

                                     

 

pGLO Transformation Lab

Purpose:

The purpose of this experiment is to transform E. Coli To be able to glow in the dark and become antibiotic resistant. 

Info:

The expression of certain traits is not only regulated by enzymes, but also by genes as well.  Enzymes usually regulate activity in the short term, while cells can regulate gene expression for the entire life cycle of many cells, making it long term.  Gene expression works in a away very similar to the way feedback inhibition in enzymes works.  It is controlled through regulatory genes and operons.  The regulatory gene controls the production of a certain molecule, but only with the help of an operon.  An operon consists of a promoter region, operon genes, and an operator.  The operator is a section of DNA that a repressor can bind to, and subsequently turn it off.  Inducible operons are ones that are normally turned off, but the presence of a certain substance can turn it on.  Repressible operons are ones that are normally on, and the presence of a certain substance can turn them off.  

In experiments where one type of foreign DNA is inserted into an organism, a plasmid is used.  A plasmid is circular bacterial DNA.  For example, the gene found in deep sea squid that makes them fluorescent, called GFP, is inserted into bacterial cell and creates a plasmid.  But, when scientists want to control an experiment where a plasmid is inserted into an organism ( in this case, bacteria) , another gene must be inserted into the plasmid.  In order to make sure that every bacterial cell being observed contains the gene for GFP, an gene for antibacterial resistance is also inserted into the plasmid. Because both the gene for GFP and antibacterial resistance are contained in the plasmid, it guarantees that only bacteria that contains the plasmid will survive for observing.

 

Methods:

 For the E. Coli to be able to glow in the dark we implanted the gene for fluorescence (Green Fluorescent Protein) into the bacteria via a plasmid. We used the plasmid pGLO that contains the gene for Green Fluorescent Protein as well as antibiotic resistance to the antibiotic ampicillin. We used 4 different agars all containing different combinations of various solutions (transformation solution, LB nutrient broth and pGLO) to test which would give us the end result of glow in the dark and antibiotic resistant E. Coli.

 

Discussion:

In this lab, we tested the transformation of E Coli. Our results showed that with the addition of pGLO to the bacteria, it was able to transform to become antibiotic resistant, and to glow. Our control plates show that without the pGLO, the bacteria doesn’t transform. Neither control plate glowed; the bacteria without antibiotic ampicillin grew, and the bacteria with ampicillin did not, exactly as we expected. The experimental plates with the pGLO added had different results: The plate with just pGLO, LB nutrient broth, and antibacterial insulin had bacterial growth, showing that the pGLO caused the bacteria to form antibiotic resistance. In the second experimental plate containing pGLO, LB nutrient broth, antibacterial insulin, and transformation solution, the bacteria not only grew, but also glowed. This shows that the pGLO caused the bacteria to form antibiotic resistance and the transformation solution caused the bacteria to glow. Our results are exactly as expected, with no growth in the control plates, and the different results of the experimental plates. We were expecting some variation because we forgot to let the solutions sit at room temperature for 10 minutes before putting them in the incubator. Our observations show that this didn’t seem to have an effect on the lab, as we got the same results anyway.  Our transformation efficiently was 1.9 x 10 to the 3, which shows that we had very high efficiency of the cells receiving the plasmids.  Overall, the expiriment was pretty successful.  

Conclusion:  In this experiment, recombinant DNA is was only viable (alive and glowing) when it contained the genes that coded for antibacterial resistance and glowing capabilities, and when arabinose was present.  In conclusion, plasmids may be inserted into bacterial DNA as a way of creating a recombinant cell, or, a cell that contains DNA from more than one organism.

 

Data:

 

Discussion:

In this lab, we tested the transformation of E Coli. Our results showed that with the addition of pGLO to the bacteria, it was able to transform to become antibiotic resistant, and to glow. Our control plates show that without the pGLO, the bacteria doesn’t transform. Neither control plate glowed; the bacteria without antibiotic ampicillin grew, and the bacteria with ampicillin did not, exactly as we expected. The experimental plates with the pGLO added had different results: The plate with just pGLO, LB nutrient broth, and antibacterial insulin had bacterial growth, showing that the pGLO caused the bacteria to form antibiotic resistance. In the second experimental plate containing pGLO, LB nutrient broth, antibacterial insulin, and transformation solution, the bacteria not only grew, but also glowed. This shows that the pGLO caused the bacteria to form antibiotic resistance and the transformation solution caused the bacteria to glow. Our results are exactly as expected, with no growth in the control plates, and the different results of the experimental plates. We were expecting some variation because we forgot to let the solutions sit at room temperature for 10 minutes before putting them in the incubator. Our observations show that this didn’t seem to have an effect on the lab, as we got the same results anyway.

Conclusion:

 In this experiment, recombinant DNA is was only viable (alive and glowing) when it contained the genes that coded for antibacterial resistance and glowing capabilities, and when arabinose was present.  In conclusion, plasmids may be inserted into bacterial DNA as a way of creating a recombinant cell, or, a cell that contains DNA from more than one organism.

References: Campbell Biology 9th edition

 

 

 

DNA Replication

Here's our review video about DNA replication!  DNA is arguably one of the most important processes in life, but can be a little confusing sometimes.  In this video, we broke replication down to make it easier to understand.

Watch and enjoy!

http://youtu.be/uCK2bopIeA0 

Cell Communication Lab

 

Purpose:

The purpose of this experiment is to examine the reproduction of yeast cells and how cellular communication plays into it.

 

Intro:

Cellular communication occurs via chemical signals that coordinate functions and respond to stimuli. Chemical signals can come from the environment, other cells in the same organism, or other organisms. Cells can communicate by direct contact, local signaling, or long distance signaling. Yeasts are unicellular fungi that can reproduce sexually or asexually. They alternate between haploid and diploid phases. During asexual reproduction, single haploid cells turn into budding haploid cells, which creates a new daughter cell. During sexual reproduction, in response to a specific chemical signal, haploids can change from an asexually reproducing cell to a gamete. There are two sexes: a-type and alpha-type. When these types mix, they grow towards each other, forming shmoos. The shmoos then fuse to create a zygote, which divides to form new yeast cells. 

 

Methods:

After placing the yeast solution (alpha type, a type and mixed) into seperate test tubes we measured using the light microscope; to find the starting amount of yeast cells. Then took measurements at different increments of time (30 minutes, 24 hours and 48 hours) to see what percent increase the yeast colony experienced periodically.

 

 

                                                      

                                                                   Mixed at 30 minutes

                                                    

 

                                                  

                                                                       Mixed at 48 hours

 

Data: 

 

 

                                                           Data for A and alpha yeast

 

 

                                                                Data for mixed yeast

Graphs:  

 

 

 

 

                                     

 

 

Discussion: 

In this lab we tested the reproduction patterns of alpha type and a type cells.  We also tested a solution of mixed alpha and a type yeast.  What we saw was this:

Alpha type yeast: Over time, the percentage of single haploid cells decreased.  At 0 minutes 77% of the cells were single haploid cells.  At 30 minutes, 76%, and at 24 hours, about 22%.  This can be attributed to the fact that there would be more formation of budding haploid cells due to the progression of time, so single haploid cell percentage would decrease.  But, we saw an increase after 48 hours, where the percentage hit 85%.  There was such a large increase I yeast concentration that the percentage of single haploid cells must have gone up.  After a certain amount of time, there must be a pattern where single haploid cells will produce budding haploid cells, and this is what happened here.  For the budding haploid cells, we saw a rapid increase.  Starting at about 23%, it increased to 24% and 77% during 30 minutes and 24 hours respectively.  But, we also saw a decrease in percentage at the 48 hour mark (14%).  

 

A type yeast: We experienced much of the same trends for the a type yeast.  Over time, single haploid cells decreased at first them increased rapidly.  It started at 89%, at 30 minutes it was at 40%, and at 24 hours it was a 15%.  But, at the 48 hour mark, it increased rapidly (85%).  For budding haploid cells, it started at 10%, increased to 60%, increased again to 84%, then decreased to 14%.  The data we had was very similar for both a and alpha type yeast, leading us to believe that they reproduce in much of the same way.

 

Mixed yeast: This sample was a combination of the a type and aha type yeast cells.  For the single haploid cells, we saw a steady decrease.  It started at a little less than 50% of the total cell count, and ended at about 29%.  This is because single haploid cells will become budding cells and eventually form asci over time, so the cell count will continually decrease.  For budding haploid cells, we observed an overall decrease as well, starting with about 17%, it gradually decreased to 7.4%.  This too, can be attributed to the fact that cells will start to form asci over time, therefore decreasing the number of free floating cells.  For the shmoos, we saw a maximum percentage of 21.8% at 30 minutes.  It then decreased to close its original percentage at 24 hours, but then increased to 17.7% at 48 hours.  For the single zygotes, we saw 7.6% at the 0 minutes. Then we saw 5.9%, 19.5%, and 11%.  For budding zygotes, the data varied by increasing, describing, and increasing once more.  The most important sample from this experiment was the percentage of asci.  It started at a relatively low percentage of 6.4%.  It ended up being at 27.4%.  This is important because as time goes by, yeast are able to communicate as a whole.  When time passes, more yeast communicate with each other,  so they are able to come in close proximity to eachother, and form asci.  

 

Basically, for a and alpha type, when time progresses, we see the overall number of yeast cells increase.  We can see that yeast has the ability to reproduce because of the increase in budding haploid cells, and therefore an increase in overall cell count (yeast cell count must increase because it is reproducing).  For the mixed type, we see an increase in the percentage of asci, which means that there was an an increase in cell communication due to the lapse of time.  Overall, the cells that denote the reproduction of yeast increase over time.  We know that yeast can't physically move because, at zero minutes, it would all conglomerate together.  Rather, it communicates by means of chemical signals.

Conclusion:

From this lab we can conclude that over time the yeast concentration increases. It is able to do so because it communicates bby means of chemical signals called pheromones. When time passes the concentration of yeast increases showing thaat it is able reproduce through chemicall signals. 

 

 

 

 

 

Plant Pigments & Photosynthesis Lab

Plant Pigments Lab

Purpose: 

The purpose of this experiment is to identify the pigmengs in chlorphyll and test how fast the pigments move up the chromatography paper.  This helps establish Rf values. 

Intro: 

Paper chromatography is used to seperate and identify pigments in cell extracts. The solvent moves up the paper because of the attraction of solvent molecules to the paper and the attraction of solvent molecules to each other. The solvent carries the pigment up the paper with it, but the pigments move up at different rates depending on how soluble they are and how much they are attracte to the paper.  Pigments in plants are used to trap light energy in order to perform photosynthesis.  The photons in light excite the electrons created from the splitting of water, and because pigments are able to harness that energy from photons, photosynthesis is able to be carried out. 

Methods:

First, we obtained a 50ml graduated calendar that contained 1cm of solvent in it.  Then, we cut a strip of filter paper and cut a point onto the end.  Next, we drew a line 1.5 cm above the tip of the point and placed a small leaf on it.  We then crushed the leaf onto the filter paper with a coin.  Next, we put the filter paper in the solvent. Once the solvent was within 1 cm of the top of the filter paper, we removed the paper and marked where each line stopped (both the pigment and solvent line).  We then measured the distance between the original and current line.  We repeated these steps for several other pigments as well.  The pigments used were Beta Carotene, Xanthophyll, Chlorophyll A, and Chlorophyll B.

Data/Graphs and Charts: 

Discussion: 

In This lab we used Chromatography paper to test the solublity of the different pigments found in spinach leaves. By placing the chromatography paper into the solvent the pigments where drawn upwards by Capillary action. Through Capillary action the solvent molecules stick to the paper and to each other. Which is quite similar to cohesion and adhesion of water molecules! When then the process was complete the solvent had left behind particles of the different pigments that were in the spinach leaf. We had 5 different lines of pigments that spread across the paper from top to bottom. From the point where we placed the spinach leaf molecules on to the paper was about 20 mm from the tip of the paper, the color of the pigment was a dark yellow color meaning that this is most likely Chorophyll B. Then the next strip of left over pigment molecules was at 40 mm and green in color. This is the band of Chlorophyll A molecules. the chlorophyll molecules are closer to the orginial pigment molecules of spinach because they are bound closer to the paper than the other pigments. The next band of pigment was at 52 mm and was yellow-green in color.This is one of the non-major pigments that are not listed in the lab or this could mean that there was a slight error in our lab. The next was Xanthophyll which is recored at 80 mm from the bottom of the paper and is a pale yellow color. Xanthophyll is higher up the paper because it is more soluble than the chlorophyll pigments, but less soluble than the Beta Caroten, since it contain oxygen and is slowed by forming hydrogen bonds with cellulose. At the top we have Beta Caroten (a yellow-orange color), it is the highest because it is very soluble  and is not slowed by the formation of hydrogen bond with cellulose, moving it up the paper the highest. For the relationship that is shown between the distance the pigment migrated from the front and from the distance solvent front migrated we had: Beta Carotene- 2.142, Xanthophyll- 1.124, Cholorophyll A- 0.571 and Chlorophyll B- 0.286. This data shows that Beta Carotene traveled the furthest from the original band of pigment where the spinach molecules were placed and supports our findings from the experiments. 

Conclusion: We concluded that Beta-Carotene moved the farthest up the chromatography sheet.  The Rf factor for Beta-Carotene was 1.  The pigment that moved the least was Chlorophyll-B.  Its Rf factor was .286.

Photosynthesis Lab 

Purpose: 

This experiment tests the hypothesis that light and chloroplasts are needed in light reactions. DPIP will take the place of NADP, which will be reduced. This will cause the DPIP to turn from blue to colorless. Based on the measurements of light transmittance, we will be able to tell which solutions have the highest rate of photosynthesis.

Intro: 

During photosynthesis, light is absorbed by leaf pigments and the electrons gain energy. Then, NADP is reduced to NADPH. ATP and NADPH are then used in carbon fixation, which turns CO2 into organic molecules.  Photosynthesis needs several things in order to be carried out.  The first thing is the presence of light.  Light excites the electrons that exist from the splitting of water, and those electrons are then able to make their way down the electron transport chain.  Another thing that is necessary for photosynthesis to occur is water.  Water is the source of electrons that help create the difference in proton gradient, which then causes ATP to be created.  Also, a plant must have a pigment that has the ability to absorb light.  The main pigment is chlorophyll-a, but many other photoreceptive pigments exist.  Carotene and chloraphyll-b are examples of other pigments.  The last thing that is needed for photosynthesis is active proteins.  Proteins make up the electron transport chain found in photosystem I and photosystem II, and there are also proteins used in carbon fixation during the Calvin cycle.  

Methods:

Day 1

 First, we prepped five different cuvettes with different solutions.  In the first cuvette, we put only the phosphate buffer, water, and chloroplasts.  In the second cuvette, we put DPIP, water, and unboiled chloroplasts, and phosphate buffer, but we placed a layer of tinfoild around it to mimic a dark environment.  In the third cuvette, we put inDPIP, water, and unboiled chloroplasts, and phosphate buffer.  In the fourth cuvette, we put in DPIP, water, and boiled chloroplasts, and phosphate buffer.  In the fifth cuvette, we put DPIP, water, phosphate buffer, but no chloroplasts.  The chloroplasts weren't put into the solution until they were ready to be put under the light.  Next, we calibrated the colorimeter with the blank cuvette (the one containing only DPIP and water) in order to establish what 0% transmittance was.  Next  we measured the reast of the cuvette in the colorimeter.  We then put the cuvette behind the lamp (with the heat recepticle between the lamp and the cuvettes), and kept time for five minutes.  Once that was complete, we placed each cuvette in the colorimeter to measure the % transmittance.  We repeated these steps for time increments of 5, 10, and 15 minutes as well.  Listed below is the contents of the solutions for each cuvette.

        

        

Day 2
We used the same methods for day 1, but instead we put 2 drops of DPIP in.

Data:

                                                                             Day 1

Day 2

Graphs and Charts:

Discussion:                

Day 1

In this experiment we were testing how chloroplasts and lights are essential for light reactions to take place. We also substituted NADP for DPIP, which is another electron acceptor. When the light hits the chloroplasts the electron levels are raised and will reduce the DPIP. This will cause the chloroplasts to change from blue to colorless, this proves that chloroplasts and light are needed because when the solution changes to colorless that means that the electron acceptors are being used by the electrons brought into the solution by light. Our control would be Sample 1, which was the blank that had no DPIP added but had the unboiled chloroplasts. In this sample we had add amount of transmittance , there was a reaction that was near 100% transmittance. Since this is the cuvette we used to calibrate the spectrometer. The second smaple was the unboiled chloroplasts without light and DPIP, with this there should be no reaction so the solution stayed blue. We placed the foil over the cuvette to block the light which stimulates the reaction. Since we had a low amount of transmittance ( about 98.5% at the end of day 1 and 71% on day 2).  This indicates that human error did occur.  The third sample of unboiled chloroplasts with light had a amount of tranmittance, this is the sample where all the necessary components are available for photosynthesis to function properly (Light, Unboiled chloroplasts & DPIP) then the solution changed to colorless. Sample 4 had boiled chloroplasts with light and DPIP, and at the end of day 1, the percent transmittance was at 94.81, and at the end of day 2, it was 66.49%. Because the chloroplasts weren't able to function due to the denaturing of the proteins involved in photosynthesis, this percentage was lower compared to the rest of the samples. The last sample 5 has no chloroplasts what so ever, meaning that there can't be a reaction because without chloroplasts the percent transmittance was nonexistant on behalf of light having to strike the chloroplasts for this reaction to take place. The research we obatined from this experiment does for sure prove that you need chloroplasts, an electron acceptor and light. 

Day 2

On day two our challenge was to somehow slow the reaction, so that our percent transmittance wasn't so high.  It jumped about 88% to about 98% percent, so tracking the increase in transmittance was more difficult.  We decided in our lab group to increase the DPIP concentration by 1 mL. By doing this we found that the change in transmittance was less drastic in the 15 minutes of testing period. This successfully translates to  slowing the reaction rate. Since we increased the electron acceptor concentration the reaction couldn't go out of control since the electron acceptors are what play a major role in controlling the reaction rate. 

Conclusion: 

Both the dark and light cuvettes (cuvettes 2 and 3) experienced the highest rate of transmittance.  Cuvette 2 had a final transmittance reading of 98.89 on day one, and 71.16 on day two. Cuvette 3 had a final transmittance reading of 98.05 on day one, and 69.70 on day two.  Although cuvette 2 did have a high transmittance rate, it was most likely caused by human error.  The rate of photosynthesis occurs slower in the dark, and eventually doesn't occur at all, so cuvette 2 must have been exposed to light accidentally.  Therefore, chloroplasts that haven't been denatured by boiling and are exposed to light will have the highest rate of photosynthesis.  

Cellular Respiration Lab

Purpose: 

In this lab, we wanted to test the whether germinated seeds and non-germinated seeds would respire, and if so, the rate at which they do so.  Also, we wanted to see how temperature impacted rate of respiration as well.  

Introduction: 

Cellular respiration is the process in which organic molecules are broken down in order to create energy for an organism.  Cell respiration occurs aerobically, which mean that oxygen is necessary for the process to occur.  Cell respiration also occurs in the mitochondria.  Before respiration begins  glucose is broken down into two molecules of pyruvate in a process called glycolosis.  If oxygen is present, then respiration will occur next.  But if oxygen isn't present, fermentation will occur.  When oxygen is present, the two pyruvate molecules are converted to AcetylCoA, which is a coenzyme.  One molecule of CO2 is released in this stage.  Next, the citric acid cycle (or Kreb's cycle) occurs.  In this stage of respiration,  2 molecules of CO2 are released as a byproduct of the breakdown of pyruvate. Two pyruvates are present, so each molecule creates one ATP and 2 molecules of CO2.  Next, the pyruvates undergo oxidative phosphorylation.  In this stage, the electrons carried by NADH (NADH collects extra electrons from molecules throughout respiration) we released onto the electron transport chain, and are subsequently carried down to increasingly more electronegative molecules.  The spillover of extra electrons in this stage fuels the transport of H+ ions across the membrane of the mitochondria, creating a large difference in energy.  When the H+ ions travel across the membrane to once again, Chemiosmosis happens.  The movement of H+ ions across the membrane to a lower concentration create energy for the synthesis of ATP.  32 to 34 ATP are created in this stage.  Overall, 2 ATP are created during glycolosis, 2 ATP are created during the citric acid cycle, and 32 to 34 ATP are created during oxidative phosphorylation.  A way to see if respiration occurs is to monitor the output of CO2 in organisms, because CO2 is emitted in both the conversion of pyruvate to AcetylCoA and the citric acid cycle.

Germination of seeds is the process in which seeds become active and able to grow.  This happens because seeds that are dehydrated intake water, and therefore, enzymes that need water to function are able to do so once again.  A germinated seed can have shoots coming off of it, and a non- germinated seed looks shriveled and water-less.

                                                 

                                                        Non-germinated seeds

                                               

                                                            Germinated seeds

Methods:
First, we collected germinated and non-germinated pea seeds.  In order to germinate the peas, we soaked them in water overnight.  Then weproceeded to test the level of CO2 emitted from several different seed types.  Here's how we did it:

Next, we separated 25 germinated, 25 non-germinated, and 25 glass beads (this was used for a control) into separate containers.

Second, we tested each of those materials for the presence of CO2 over a ten minute time span.

Third, we put the 25 germinated peas into ice-cold water for approximately ten minutes, in order of I test the affect of tempurature of germinated peas on levels of CO2 output.  We then tested the cold peas for presence of CO2 as well.

Lastly, we created graphs showing the CO2 output over ten minutes.

Data:

       

Graphs and Charts:

                                                          CO2 Release vs. Time Graph

    

Discussion: 

In this lab, we tested cellular respiration in different types of seeds. We did this by calculating the amount of CO2 emitted by the seeds, since CO2 is a product of cellular respiration. Our group tested peas that were germinated at room temperature, peas that were germinated in ice water, and dormant seeds. We also used glass balls as a control group. Our results showed that at 0.80 ppm/s, peas germinated at room temperature emitted the most carbon dioxide, showing that they have the highest rate of cellular respiration. The germinated peas in ice water had a lower rate, at 0.71 ppm/s, showing that cellular respirations is still occurring in lower temperatures, just at a lower rate. The dormant peas were drastically lower, with a rate of 0.15 ppm/s. Our control group of glass beads had a rate of 0.10 ppm/s, showing that a small amount of carbon dioxide could have been coming from other places, such as the water trapped in the bottle, it could've been leaking into the bottle, or it could have been trapped in the bottle from the previous test. Overall, our results showed that a germinated cell has a higher rate of cellular respiration than a non-germinated seed, as well as there is a direct relationship between temperature and respiration: as temperate goes up, so does respiration; as temperature goes down, so does respiration. 

Conclusion: From this experiment we can conclude that the non-germinated seeds do not respire, as shown by the very little amount of CO2 that was being given off by the non-germinated seeds. As for the germinated seeds we found that when the seeds were at room temperature that they respired at the highest rate.  The cold seeds had a lower amount of CO2 then the room temperature seeds. In summary when the temperature is higher the respiration of CO2 is higher than those of seeds in colder environments and that non-germinated seeds do not respire.

References: 

http://plantsinmotion.bio.indiana.edu/plantmotion/earlygrowth/germination/germ.html

Campbell Biology, 9th edition