The methods of the moment

The investigation that I’m doing requires the techniques of extraction, amplification and electrophoresis.
Many if these procedures take some time because there are many steps on to making the procedure. They all turn up being a BIG chain of events that end up with electrophoresis and by this (mainly) studying the DNA that has been cleaned, amplified by PCR, separated by restriction enzymes and finally observed in the gel of the electrophoresis. With the reading of the gel I can determine if the restriction enzymes that I use produce a proper Bio-marker or do I need to find different ones to have the job done.

Extraction

In basic terms, this process that haves many steps basically isolates and cleans the DNA of the bacteria that that I wish to study.  Common chemicals used are Ethanol and Chloroform are basically used for the cleaning of fats and any other residue that isn’t DNA. And the Lysis Buffer solution fragmentates the bacteria so the DNA is poured out. RESULT: CLEAN DNA.

PCR

Is the step in which we make many replicates of the DNA that I am interested with. Useful because makes DNA more visible in electrophoresis and if some DNA is lost during procedures y have many copies to replace it. A common name in this procedure is the primer which marks the part of sequenced bases that I want the “coping” to start from. This step is commonly called the amplification of the DNA.

Electrophoresis

This is the step that brings out the “readable” data. Charge repulsion is the used rule for this method. The DNA carries a negative charge and so do electrons. The method basically is to carry a current so the DNA (repulsed by the negative charge) is repulsed. The heavier strands of DNA stay behind meanwhile the lighter travel farther on. This strands are divided if we “cut” the DNA with a restriction enzyme. If this is not done, a single and heavy strand of DNA will be the result. We can know how many base pairs does the strand have because a “ladder” ( its like a legend) tells us how many base pair we may have. It may sound like a very complicated process but, once you do it for the first time, it will be very easy to understand.

 BIO-BLOG COMMENTS:

Ive just submited the comments and I have seen that some are having the same problems that I am confronting. I notices that one of the students is going to do the same procedure that I am. I think that it is a good thing that the students are connecting because we can healp and support each other. Many nice blogs and I hpe to see many more.