+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Pressure-based alkaline lysis with immunocapture, a method for enhanced recovery in differential extraction



Pressure-based alkaline lysis with immunocapture, a method for enhanced recovery in differential extraction



Electrophoresis 38(21): 2777-2785



A pressure-based protocol for differential extraction has been optimized to provide a rapid and selective alternative to conventional differential extraction techniques with the advantage of an increased recovery of genomic DNA. The protocol involves treating cotton swabs containing mixtures of sperm and vaginal epithelial cells with two sets of pressure treatment using a Barocycler® NEP 2320 in alkaline conditions. This first step quantitatively and selectively removes female epithelial cells. The cotton swab is removed and further treated with alkali at 95°C for the removal of sperm cell DNA. The resultant solution provides a clean male profile at a 20:1 cell ratio. Furthermore, the inclusion of a pretreatment step involving immunomagnetic cell capture of the female cells, permits nearly complete isolation of male sperm cells at cell ratios of up to 200:1 female to male cells.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 060125718

Download citation: RISBibTeXText

PMID: 28714088

DOI: 10.1002/elps.201700139


Related references

The application of alkaline lysis and pressure cycling technology in the differential extraction of DNA from sperm and epithelial cells recovered from cotton swabs. Analytical and Bioanalytical Chemistry 407(23): 6975-6984, 2015

Development of a rapid, 96-well alkaline based differential DNA extraction method for sexual assault evidence. Forensic Science International. Genetics 6(1): 1-16, 2012

A rapid method for authentication of Buffalo (Bubalus bubalis) meat by Alkaline Lysis method of DNA extraction and species specific polymerase chain reaction. Journal of Food Science and Technology 50(1): 141-146, 2013

Evaluation of an alkaline lysis method for the extraction of DNA from whole blood and forensic stains for STR analysis. Journal of Forensic Sciences 45(3): 669-673, 2000

Validation of a fast and low-cost alkaline lysis method for gDNA extraction in a pharmacogenetic context. Cancer ChemoTherapy and Pharmacology 75(5): 1095-1098, 2015

Extraction of sperm DNA from mixed stain by the modified differential lysis method combined with silicon bead method. Fa Yi Xue Za Zhi 30(1): 50, 2014

Deparaffinization and lysis by hydrothermal pressure (pressure cooking) coupled with chaotropic salt column purification: a rapid and efficient method of DNA extraction from formalin-fixed paraffin-embedded tissue. Diagnostic Molecular Pathology 22(1): 52-58, 2013

Material modifications for the alkaline differential extraction method for sexual assault evidence. Forensic Science International. Genetics 7(4): E104-E105, 2013

Enhanced recovery and purification of P(3HB-co-3HHx) from recombinant Cupriavidus necator using alkaline digestion method. Applied Biochemistry and Biotechnology 167(3): 524-535, 2012

Enhanced recovery of chlorophenols from surface waters using polymer based extraction cartridges. Fresenius' Journal Of Analytical Chemistry. 360(1): 90-94,., 1998

Differential inactivation of three bacteriophages by acid and alkaline pH used in the membrane adsorption-elution method of virus recovery. Canadian Journal of Microbiology 26(12): 1403-1407, 1980

Obtaining an additive based on resinous bark for the alkaline pulping of beech wood. II. Process for enhanced extraction of additives. Celuloza si Hirtie 39(2-3): 65-69, 1990

Single-cell copy number analysis of prostate cancer cells captured with geometrically enhanced differential immunocapture microdevices. Analytical Chemistry 86(22): 11013-7, 2014

Sequential alkaline saponification/acid hydrolysis/esterification: A one-tube method with enhanced recovery of both cyclopropane and hydroxylated fatty acids. Journal of Microbiological Methods 18(1): 21-32, 1993

Optimizing alkaline lysis for DNA plasmid recovery. Biotechnology and Applied Biochemistry 37(Pt 3): 235-244, 2003