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Rosengarten, RD Rafael D, Beltran, PR Pamela R, Shaulsky, Gad, (2015) ' A deep coverage Dictyostelium discoideum genomic DNA library replicates stably in Escherichia coli. ' Genomics

 

Abstract:The natural history of the amoeba Dictyostelium discoideum has inspired scientific inquiry for seventy-five years. A genetically tractable haploid eukaryote, D. discoideum appeals as a laboratory model as well. However, certain rote molecular genetic tasks, such as PCR and cloning, are difficult due to the AT-richness and low complexity of its genome. Here we report on the construction of a ~20 fold coverage D. discoideum genomic library in Escherichia coli, cloning 4-10 kilobase partial restriction fragments into a linear vector. End-sequencing indicates that most clones map to the six chromosomes in an unbiased distribution. Over 70% of these clones contain at least one complete open reading frame. We demonstrate that individual clones and library composition are stable over multiple replication cycles. Our library will enable numerous molecular biological applications and the completion of additional species' genome sequences, and suggests a path towards the long-elusive goal of genetic complementation.
Status:	aheadofprint	Type:	Journal article	Source:	PUBMED	PubMed ID:	26028264

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	Genes addressed in this paper
	17S_rRNA-1	17S_rRNA-2	DDB_G0290575	H3a	cprD	csaA	dnase2	pks23
Topics in this paper
Genome-wide Analysis	X	X	X	X	X	X	X	X
DNA/RNA Sequence Features	X	X	X	X	X	X	X	X
Techniques and Reagents	X	X	X	X	X	X	X	X

	Genes addressed in this paper
	rabK1	rabK3	rabL	rabM	rabN1	spiA	srfA
Topics in this paper
Genome-wide Analysis	X	X	X	X	X	X	X
DNA/RNA Sequence Features	X	X	X	X	X	X	X
Techniques and Reagents	X	X	X	X	X	X	X