Seamless Ligation Cloning Extract (SLiCE) Cloning Method - DNA Cloning and Assembly Methods - page 230

Fig. 1 SLiCE cloning of a single DNA fragment. ( a ) SLiCE cloning without fl anking
heterologous sequences. ( b ) SLiCE cloning with fl anking heterologous sequences
(Reproduced from [ 4 ], with permission from Oxford University Press)
homologies (15-52 bp) in bacterial cell extracts termed SLiCE
( S eamless Li gation C loning E xtract) [ 4 ]. SLiCE allows for effi cient
restriction site-independent cloning of DNA fragments generated
by restriction digestion or PCR amplifi cation into linearized vec-
tors. In addition, SLiCE does not require the use of enzymes for
the modifi cation of vector and insert end sequences (such as
Klenow or T4 DNA polymerase) or ligases.
The SLiCE method [ 4 ] can be used for virtually any type of
cloning approach such as the cloning of a single DNA fragment
with or without fl anking heterologous sequences ( see Fig. 1 ), the
cloning of multiple DNA fragments in one step ( see Fig. 2 ), and
the directional subcloning of large DNA fragments from bacterial
artifi cial chromosomes (BACs) ( see Fig. 3 ).
The SLiCE method is based on bacterial extracts that can be
derived from a variety of common RecA − E. coli laboratory strains
such as DH10B and JM109 [ 4 ]. In vivo homologous recombina-
tion in E. coli can be facilitated by three different recombination
pathways: The RecA-dependent pathway, an RecA-independent
pathway of unknown nature and an RecA-independent pathway
that utilizes prophage Red/ET recombination systems [ 5 - 10 ].