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In vitro packaging of lambda and cosmid DNA.
High efficiency, restriction-deficient in vitro packaging extracts for bacteriophage lambda DNA using a new E.coli lysogen.
Guo, P., Erickson, S. & Anderson, D. A small viral RNA is required for in vitro packaging of bacteriophage phi 29 DNA.
A 15-kilobase pair EcoRI chick DNA fragment, containing both the termination codon UGA and the 5'-portion of the structural ovomucoid gene, has been cloned in lambda phage Charon 4A by in vitro packaging.
Nevertheless, T7 and λ systems rely on in vitro packaging systems that are quite labile and have limitations for the construction of libraries with large number of variants with a limit up to 107 to 108 variants.
Plasmid pGL3-control (Promega cat # E1611), which carries the luc reporter gene expressed from the early SV40 promoter, was used for the in vitro packaging experiments.
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Unlike other in-vitro packaging systems, no viral genetic material or packaging signal sequence is required to form these pseudovirions.
The in vitro packaged virions produce distinctly blue plaques when plated on a suitable host.
The ligated DNA products were then in vitro packaged using Gigapack III packaging extract (Stratagene, USA).
The recombinant cosmid were in vitro packaged and transfected into E. coli XL1-blue MRF' strain.
Insert-DNA was ligated in the pCC1-Fos™ vector, constructs were in vitro packaged into phage particles and transfected into Escherichia coli EPI 300™-T1R.
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