DNA sequence detection of nucleated red blood cells
First Claim
1. A method for isolating nucleic acid from a chicken comprising:
- obtaining a blood sample comprising nucleated blood cells from a bird;
adding to a container, the blood sample and a plasma membrane lysis buffer wherein the container binds a precipitated nucleic acid;
centrifuging the container to yield a supernatant and a pellet;
removing the supernatant from the container;
adding to the container a nucleic acid release lysis buffer;
incubating the container such that nucleic acid is released;
precipitating the nucleic acid samples in the container producing a supernatant;
removing the supernatant; and
dissolving the nucleic acid in a solvent, thereby isolating nucleic acid from a bird.
2 Assignments
0 Petitions
Accused Products
Abstract
Genetic modification or selection of avians requires that large numbers of birds be genetically analyzed for sequences of interest. Typically, DNA is extracted on an individual basis from samples taken from the birds. Current methods of DNA extraction extract the DNA from blood or other tissues using tedious and time-consuming procedures. The present invention provides a high throughput screening assay for detecting a genetic sequence in multiple samples. The assay further provides a DNA extraction method that allows DNA to be extracted rapidly from multiple avian samples, such as red blood cells. The extraction method is extremely reliable and does not require that each sample be quantitated post-extraction. The extracted DNA can be used for a variety of genetic assays, including a high throughput screening assay to identify insertion of a transgene. The present invention is particularly useful for extracting DNA from nucleated RBCs. Therefore, the method can be applied towards genetic analysis of avians, fish, reptiles and amphibians.
-
Citations
15 Claims
-
1. A method for isolating nucleic acid from a chicken comprising:
-
obtaining a blood sample comprising nucleated blood cells from a bird;
adding to a container, the blood sample and a plasma membrane lysis buffer wherein the container binds a precipitated nucleic acid;
centrifuging the container to yield a supernatant and a pellet;
removing the supernatant from the container;
adding to the container a nucleic acid release lysis buffer;
incubating the container such that nucleic acid is released;
precipitating the nucleic acid samples in the container producing a supernatant;
removing the supernatant; and
dissolving the nucleic acid in a solvent, thereby isolating nucleic acid from a bird. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
-
Specification