A High School Student needs your help with RNA extraction!

[emilyjo10]

Hi! My name is Emily, and I am a high school senior in my Aquatic Biology/Scientific Research Honors class. For our final project, we have to write a scientific paper on anything to do with aquatic habitats. (water, fish, streams, etc.)

I am very interested in Genetics, so I was hoping to do my research on something to do with it. My teacher is allowing me access to her RNA Extraction equipment. However, she has never had a successful extraction. I would really like to maybe compare RNA of 2 different organisms from the same species.

 

The Questions:

1. Is there a specific aquatic organism's RNA that would be the best/easiest to extract?

2. How should a high school student go about figuring out this complicated process?

3. What are some more ideas/experiments I could do with the results of the RNA extraction, if it works?

 

Any other advice would be greatly appriciated!

THANK YOU!

 

Emily

[CharonY]

1) it depends on what specimen you can easily get and what equipment you got. Essentially it boils down to the methods you got which allows a proper homogenization and of your sample and effective lysis of the cells (e.g. bead beater or something similar, liquid nitrogen etc.).

2) if you use a kit, read the manual carefully. RNA extraction is in principle fairly easy, but you have to keep everything clean (ie free of RNAses). If you are not using a kit, I would recommend the book Molecular cloning (Sambrook et al.) it has a very nice chapter regarding RNA extraction protocols. Obviously, inform yourself before start the actual experiment.

3) Depends really on what you used as specimen and what further equipment you got. Quite obviously you cannot expect to generate important data by a simple experiment. But things to consider are, e.g. QPCR to measure abundance of a given RNA (provided some sequences of the specimen in question are known), amplify and clone rRNA (or rather the reverse transcribed cDNA) for sequencing and phylogenetic analyzes, etc. But I reckon that on the highschool level a) getting RNA at all is pretty good already. A simple thing you could do is check your RNA for DNA impurities.

[emilyjo10]

Thank you so much for your advice!

This is already helping me so much.

Thankfully, I am using a mirVana miRNA Isolation Kit from Applied Biosystems.

I am carefully reading and taking notes over the manual now.

Thank you again!

 

Emily