A WEB BASED INTERFACE FINDING SUB-CLONING STRATEGIES, IN-FRAME DELETIONS
AND FRAMESHIFTS USING RESTRICTION ENZYMES AND DNA POLYMERASES.
LINDENBAUM Pierre (1998) Bioinformatics/CABIOS Vol.14;5,1998 pp465-466
What is CloneIt ?
The use of plasmid vectors in cloning strategies is the basis of any
molecular biologist repertoire. Efficiency is essential and requires the
use of the most appropriate restriction enzymes choices for the desired
application. Selection of those enzymes remains a skill. Several points
that merit consideration include (1) the enzymes characteristics, (2)
location of the restriction sites within the sequence, (3) complementarity
of protuding ends, (4) possible self ligation, (5) use of modifying DNA
polymerase generating blunt ends, (6) in frame cloning constraints,(7) use
of partial digestions and (8) the creation of a stop codon at the ligation
site. This exercise is labor intensive, even with the help of the
classical DNA analysis softwares. These programs typically facilitate
localizing restriction sites in a plasmid sequence; however it is
impossible to examine all restriction sites and /or combinations then, a
simple cloning strategy may be over-looked as a result of incomplete or
CloneIt Online was created in order to quickly find cloning
strategies (including sub-cloning, in-frame deletions and frameshifts)
while controlling the problems described above.
CloneIt Online is available at the following URL: