page 2 of 7humans; therefore, the nucleotide sequences of any two humans is roughly 99.9%identical. The polymorphism rate may be substantially higher in other species.Additionally, bases possessing the highest polymorphism rates are typically leastimportant, often falling in non-functional DNA where mutations in the nucleotidesequence have least impact on the organism's survival. Thus, similarity is preserved.
3Tools of DNA Sequencing
: small circular pieces of DNA.
Using restriction digest enzymes, the sample DNA is cleaved into shorter (~ 10
bases) fragments. These restriction enzymes only cleave at specific recognitionsequences in the DNA. The vector is then cleaved using the same restrictionenzymes, allowing the DNA fragments (inserts) to incorporate into the vector.
VectorSize of Insert
Plasmid2,000-10,000(can control size)Cosmid40,000BAC (Bacterial Artificial Chromosome)70,000-300,000YAC (Yeast Artificial Chromosome)> 300,000(not used much recently)
Genomic DNA to be sequencedRestriction enzyme digestDNA fragmentsVector Circular genome(bacterium, plasmid)Knownlocation(restrictionsite)