منابع مشابه
Efficient random subcloning of DNA sheared in a recirculating point-sink flow system.
Based on a high-performance liquid chromatographic pump, we have built a device that allows recirculation of DNA through a 63-microm orifice with ensuing fractionation to a minimum fragment size of approximately 300 base pairs. Residence time of the DNA fragments in the converging flow created by a sudden contraction was found to be sufficiently long to allow extension of the DNA molecules into...
متن کاملA subcloning strategy for DNA sequence analysis.
We describe here a new strategy of fragment preparation for sequencing procedures using endlabelled DNA fragments as substrates (2,3) which is directly applicable to DNA fragments cloned into the Pst I site of pBR322, or in modified form, to inserts into the BamH I or Sal I site of the same plasmid. Ordered sets of subclones of predetermined overlap are are generated. These can be sequenced dir...
متن کاملSubcloning of the Histone DNA Sequences
The purification and characterization of histone mRNA is a relatively straightforward procedure if conducted with the RNA from early cleavage stage sea urchin embryos [1 -6 ] where histone mRNA is the most abundant mRNA component found in small polysomes [1, 2]. Consequently this has led to an extensive characterization of the different subtypes of histone mRNA [4 -9 ] as well as to their use a...
متن کاملIsolation and subcloning of large fragments from BACs and PACs.
Bacterial artificial chromosome (BAC) vectors and P1-derived artificial chromosome (PAC) vectors are widely used for structural and functional analysis of genomic DNA in vitro and in vivo. The vectors can harbor fragments up to 300 kb in size and show high stability even with alphoid repetitive inserts (7). For structural analysis, they have great advantages because of their stability and low c...
متن کاملEfficient DNA subcloning through selective restriction endonuclease digestion.
Described here is a selective restriction endonuclease digestion method that eliminates the electrophoresis step that is usually used during the subcloning of new DNA sequences into typical E. coli-based plasmids. The method increases yield while decreasing laboratory resource and time utilization. By using donor and acceptor sequences that contain unique restriction sites found only outside of...
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ژورنال
عنوان ژورنال: Genome Research
سال: 1995
ISSN: 1088-9051
DOI: 10.1101/gr.5.5.464