ArticleNucleic acids research1997
A novel method for producing partial restriction digestion of DNA fragments by PCR with 5-methyl-CTP.
Article in Nucleic acids research, 1997. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed.
- Microfluidic Continuous Flow DNA Fragmentation based on a Vibrating Sharp-tip.Microfluidics and nanofluidics · 2022Article
- The enhancement of DNA fragmentation in a bench top ultrasonic water bath with needle-induced air bubbles: Simulation and experimental investigation.Biomicrofluidics · 2022Article
- High Efficiency Hydrodynamic DNA Fragmentation in a Bubbling System.Scientific reports · 2017Article
- Molecular tandem repeat strategy for elucidating mechanical properties of high-strength proteins.Proceedings of the National Academy of Sciences of the United States of America · 2016Article
- A simple method for semi-random DNA amplicon fragmentation using the methylation-dependent restriction enzyme MspJI.BMC biotechnology · 2015Article
- Cavitation Enhancing Nanodroplets Mediate Efficient DNA Fragmentation in a Bench Top Ultrasonic Water Bath.PloS one · 2015Article
- A device for automated hydrodynamic shearing of genomic DNA.BioTechniques · 2009Article
- Identification and sequence analysis of a 27-kilobase chromosomal fragment containing a Salmonella pathogenicity island located at 92 minutes on the chromosome map of Salmonella enterica serovar typhimurium LT2.Infection and immunity · 1998Article
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3 authors.
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Abstract
Partial digestion of DNA fragments is a standard procedure for subcloning analysis and for generating restriction maps. We have developed a novel method to generate a partial digestion for any DNA fragment that can be amplified by PCR. The method involves the incorporation of 5-methyl-dCTP into the PCR product to protect most of the restriction sites. As a result, complete digestion of the modified PCR products with a 5-methyl-dCTP-sensitive enzyme will produce an array of restriction fragments equivalent to a partial restriction enzyme digestion reaction done on unmethylated PCR products. This method reduces the time and material needed to produce partially-digested DNA fragments by traditional methods. Furthermore, using fluorescein-labeled primers in the reaction, we were able to detect the fluorescein-labeled end fragments resulting from the enzyme digestion using a fluorimager or anti-fluorescein-AP antibody and thus determine the restriction maps.
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