Abstract
One of the obstacles encountered when extracting DNA from a large number of samples is the cumbersome method of deproteinizing cell digests with the hazardous organic solvents phenol and isochloroform. A rapid, safe and inexpensive method was developed to simplify the deproteinization procedure. This method involves salting out of the cellular proteins by dehydration and precipitation with a saturated NaCl solution. Buffy coats of nucleated cells obtained from anticoagulated blood (ACD or EDTA) were resuspended with 3 ml of nuclei lysis buffer. The DNA obtained from this simple technique yielded quantities comparable to those obtained from phenol-chloroform extractions. This procedure has been used in our laboratory on several thousand blood samples for parentage, population and forensic studies. This technique is used with out nonisotopic hybridization procedures rendering the entire process of RFLP analysis free of toxic materials.
Citation Formats
Miller, S A, Dykes, D D, and Polesky, H F.
Simple salting out procedure for extracting DNA from human nucleated cells.
United Kingdom: N. p.,
1988.
Web.
doi:10.1093/nar/16.3.1215.
Miller, S A, Dykes, D D, & Polesky, H F.
Simple salting out procedure for extracting DNA from human nucleated cells.
United Kingdom.
https://doi.org/10.1093/nar/16.3.1215
Miller, S A, Dykes, D D, and Polesky, H F.
1988.
"Simple salting out procedure for extracting DNA from human nucleated cells."
United Kingdom.
https://doi.org/10.1093/nar/16.3.1215.
@misc{etde_6998585,
title = {Simple salting out procedure for extracting DNA from human nucleated cells}
author = {Miller, S A, Dykes, D D, and Polesky, H F}
abstractNote = {One of the obstacles encountered when extracting DNA from a large number of samples is the cumbersome method of deproteinizing cell digests with the hazardous organic solvents phenol and isochloroform. A rapid, safe and inexpensive method was developed to simplify the deproteinization procedure. This method involves salting out of the cellular proteins by dehydration and precipitation with a saturated NaCl solution. Buffy coats of nucleated cells obtained from anticoagulated blood (ACD or EDTA) were resuspended with 3 ml of nuclei lysis buffer. The DNA obtained from this simple technique yielded quantities comparable to those obtained from phenol-chloroform extractions. This procedure has been used in our laboratory on several thousand blood samples for parentage, population and forensic studies. This technique is used with out nonisotopic hybridization procedures rendering the entire process of RFLP analysis free of toxic materials.}
doi = {10.1093/nar/16.3.1215}
journal = []
volume = {16:3}
journal type = {AC}
place = {United Kingdom}
year = {1988}
month = {Feb}
}
title = {Simple salting out procedure for extracting DNA from human nucleated cells}
author = {Miller, S A, Dykes, D D, and Polesky, H F}
abstractNote = {One of the obstacles encountered when extracting DNA from a large number of samples is the cumbersome method of deproteinizing cell digests with the hazardous organic solvents phenol and isochloroform. A rapid, safe and inexpensive method was developed to simplify the deproteinization procedure. This method involves salting out of the cellular proteins by dehydration and precipitation with a saturated NaCl solution. Buffy coats of nucleated cells obtained from anticoagulated blood (ACD or EDTA) were resuspended with 3 ml of nuclei lysis buffer. The DNA obtained from this simple technique yielded quantities comparable to those obtained from phenol-chloroform extractions. This procedure has been used in our laboratory on several thousand blood samples for parentage, population and forensic studies. This technique is used with out nonisotopic hybridization procedures rendering the entire process of RFLP analysis free of toxic materials.}
doi = {10.1093/nar/16.3.1215}
journal = []
volume = {16:3}
journal type = {AC}
place = {United Kingdom}
year = {1988}
month = {Feb}
}