Blog 4: 10 Points to Know About Biotechnology

Vector Cloning

-Vector cloning requires plasmids, restriction enzymes, ligase, and a vector (bacteria).
-"Sticky ends" are single strand sequences at the ends of the cut pieces that matches with other sticky ends made by the same restriction enzyme.

Polymerase Chain Reaction

-Successive cycles of heating and cooling can cause DNA to separate, have primers bind on, elongated by Taq polymerase and the cycle repeats.
-By the end of the third cycle, the first desired sequences are produced. By the end of the 20th cycle, over a million of the desired sequence is produced.

Gel Electrophoresis

-DNA that has been treated by restriction enzymes can be sorted into their respective lengths.
-Smaller fragments move through the gel to the positively-charged terminal (DNA has a negative charge) while larger fragments remain closer to the starting point.

DNA Sequencing
-DNA is replicated in solutions containing a mix of dideoxyribonucleotides which terminate elongation at various points due to the lack of 3' oxygen.
-By randomization, fragments of many different lengths are produce which all indicate the position of a corresponding nucleotide of the dideoxyribonucleotides.

Restriction Fragment Length Polymorphisms
-DNA from different individuals can have variation in restriction sites when treated with restriction enzymes.
-Their analysis is useful for detecting mutations, assessing risk for genetic disorders, and forensic science.