Rohith Kaliyur Properties of Algae Block 3 Individual Procedure Design Process 1

In this post I am primarily going to describe one of the procedures that I have designed, and subsequently going to mention any further inquiries or procedures that I am looking into. The procedure will not be super detailed, however, it will be fully operational, and if finalized on, 100% useful. The procedure I will be describing is for analysis of ribosomal and DNA characteristics in algae. This is an extremely difficult process to carry out and there is no guarantee that it will even work. Nonetheless, I have decided to integrate it as the analysis is seemingly very attractive.

• Procedure for Performing Gel Electrophoresis in Algae:Obtain the following materials: Electrophoresis chamber, gel form and comb, electrophoresis power supply, agar gel (preparation for this is not described but is also a process), salt solution, algal strains to use as samples, sample loading device, masking tape.

• Dissolve the agar, cool the solution, and pour the gel.

Combine agar and water. Bring the mixture to a boil and heat until the agar is dissolved

Cool the agar until you can comfortably touch the flask.

Place tape across the ends of the gel form (if needed) and place the comb in the form.

Pour cooled agar into the form; the bottom 1/3-1/2 of the comb should be immersed.

Immediately rinse and fill the agar flask with hot water to dissolve any remaining agar.

When the agar has solidified, carefully remove the comb.

Remove the tape (if used) from the ends of the gel form.

•  Load samples in the wells with the gel

Make a written record of which sample you will load in each well of the gel.

Place the gel form on a black or dark surface to help you see the wells in the agar.

You may find it helpful to only load samples in every other well.

Be careful to not puncture the bottoms of the wells 

Place the gel in the electrophoresis chamber with the wells closest to the negative (black) electrode

Prepare the salt solution and add it to the chamber.

Add salt to tap water and swirl it to dissolve.

Fill each half of the chamber, adding solution until it is close to the top of the gel. Then gently

flood the gel from the end opposite the wells to minimize sample diffusion 

• Place the lid on the chamber and connect the electrode leads to the power supply 
• Turn on the power supply and adjust the voltage to 50-100 volts
• Run the gel for 5-10 minutes

Ultimately, at the end of this procedure, a particular light differential should be observed. This can be translated into particular RNA and DNA sequences that will better the global understanding of algae.
My inquiries and further explorations involve altering lipid concentrations within algae, simulating glycolysis within algae, or trying to manipulate the photosynthesis of algae in some way or form. It is very possible that I choose not to implement any of these ideas.

Rohith Kaliyur Properties of Algae Block 3 Individual Research Process 4

This will be my last blog post in regards to the individual research I have done on my topic. I am slowly transitioning from the research phase of the project to the procedural part of the project. Although the amount of information that I have obtained in addition to my previous information has been relatively limited in terms of algal information, it has been vastly abundant in terms of other scientific information. This is due to the fact that I have allocated a significant amount of my time towards researching different scientific processes and procedures utilized by other scientists. I believe that this will give me an even greater insight towards what to do for the upcoming phase of my experimentation. One of the procedures I have been looking at in particular involves intentionally not nourishing certain algal cultures, similar to a play in Hamlet in which the king is poisoned by a seeker for the throne. After this phase, a period of observation of these algal cultures will occur, trying to predict how the respective algae will perform given alternate simulated environments. In my upcoming blog entry I am most likely going to post a personal procedure as to a particular method to obtain a particular algal property. It will most likely not be too detailed, and may not even be utilized. It is more for the brainstorming process. A lot of the procedures that I throw out there, I may realize later that I lack the funding or materials to implement. In terms of more algal properties, I have mainly been focusing on lipid production and photosynthesis. At the moment, I am trying to concur as to whether algae has more plantae or animalia features, as this will greatly aid in designing certain procedures for the overall experimentation.

At the moment here are some diagrams of certain experiments I have been looking into somewhat utilizing. These are more for the brainstorming process, however, they could very well become useful.
Diagram 1
Diagram 2
Diagram 3
Diagram 4
Diagram 5

These are just some of the experimental ideas I have been looking into, I do have more that I am researching. However, these provide a general idea of the direction I am headed.

Rohith Kaliyur Properties of Algae Block 3 Individual Research Process 3

Several major milestones were reached. First, I completed my respective publication. This publication was soaking an enormous amount of my time, and I am happy to have finally completed it. Although I am not 100% thorough with all of the expressed topics, I feel that this novel has educated me enough to allow me to adequately conduct personal research regarding a substantial topic related to algae to hopefully advance the field. Second, I narrowed down my choices as to which species of algae to use, the green algae. This is because this type of algae is vastly abundant in the environment I am in, and will thus be easy to either obtain or purchase.

Here are a few vital facts about this interesting chlorophycae:

1)They are abundant in fresh water.

2)They can occur as single cells or colonies.

3)Their main storage compound is starch, although oils can be produced under certain conditions.

I believe this algae is the evolutonary progenitor of modern plants.

I have decided to elaborate on green algae anatomy, here is a pictorial image that serves as a very basic model.

Green Algae Anatomy

In condensed form algae is similar to numerous prokaryotic organisms, as it has the ability to be unicellular. This is what is displayed in the diagram, however, in reality the anatomy varies between prokaryotic and eukaryotic algae. Although this seemingly makes algae tougher to truly understand, the overall pattern gives rise to certain similarities that allow for better exploration of its properties.

There is one more aspect I would like to touch on in this blog entry. Numerous publications and sources that I have read have impacted algal properties in many different ways to more efficient biofuels. Although this seems like a wise allocation of resources for major organizations and universities, I will not indulge in this as I simply do not have the available resources to do so. For the most part I must begin to focus an enormous amount of this project remaining towards closing out my research and beginning to design my own procedures for testing for certain properties of algae.

Rohith Kaliyur Properties of Algae Block 3 Individual Research Process 2

This post is mostly an update as most of my findings will be unveiled in my following post. I am still immersed in my particular publication, learning new and interesting things I had never before conceived were possible related to algae. That being stated, there is still much work to be done. The narrowing down to either research of multiple species of algae or a single species is still a resonating question. Despite this uncertainty, I have learned a significant amount from numerous other experiments other prominent individuals have conducted regarding algae. One example that I am currently reading about in various sources is measuring lipid concentrations of algae in desert climates. The scientist, who is anonymous in the sources, records the utilization of obtained algal samples in combination with previous research. He then proceeded to test out the algae in alternate environments and subsequently observing any changes in lipid concentrations of the algae. Although this in itself is a somewhat complicated experiment, it helped in my personal comprehension of certain processes such as how micro-algae store CO2 as lipids. Some other basic characteristics about algae are that they are in fact eukaryotic organisms. This means that they are multicellular organisms for the most part. There are unicellular algal cultures out there in rivers and streams, yet, these cultures are the minority. However, their behavior is similar to that of plants in that they carry out photosynthesis. Despite this similarity, algae do not have roots, stems, or leaves. In addition, there are 2 main types of algae, macroalgae ie seeweed, and microalgae ie the golden algae. To elaborate on my previous blog post, the primary types of algae are the diatoms (Bacillariophycae), the green algae (Chlorophyceae), the blue-green algae (Cyanophycaeae), and the golden algae (Chrysiohyceae). All of these algal types grow in different environments and deciding which to use will certainly be a challenge. For this post I won't delve too deep into the whole anatomy of algae at the moment, however, this will be done in the following post. I will only describe the anatomy of the respective species that I choose. My goals for this project are to make way with the actual experimentation as that will be the most interesting part, and to consolidate on certain properties of algae to look test for. The procedural process is significantly more difficult than I originally assumed, as it requires significant mental investment.