266
LUIZ F. ONUCHIC AND GREGORY G. GERMINO
walks (Figure 2). This problern can be overcome by using multiple libraries
that are prepared from DNA cleaved with different endonucleases.
Another majorproblern associated with one-enzyme complete digest libraries isthat they lack restriction fragments of the original source material
that arenot within the acceptable cloning Iimits of the vector (Figure 2).
Complete restriction enzyme digestion of complex genomes typically yields
fragments that range greatly in size. The proportion of the total that will be
I : Endonuclease A restriction sites
: Clonable Fragments
: Unclonable Fragments
Double-stranded genomic DNA
Fragments produced by complete digestion
Fragments produced by partial digestion
Fig. 2. Partially and completely digested libraries
LUIZ F. ONUCHIC AND GREGORY G. GERMINO
walks (Figure 2). This problern can be overcome by using multiple libraries
that are prepared from DNA cleaved with different endonucleases.
Another majorproblern associated with one-enzyme complete digest libraries isthat they lack restriction fragments of the original source material
that arenot within the acceptable cloning Iimits of the vector (Figure 2).
Complete restriction enzyme digestion of complex genomes typically yields
fragments that range greatly in size. The proportion of the total that will be
I : Endonuclease A restriction sites
: Clonable Fragments
: Unclonable Fragments
Double-stranded genomic DNA
Fragments produced by complete digestion
Fragments produced by partial digestion
Fig. 2. Partially and completely digested libraries