HIGHLY MULTIPLEXED GENOTYPING USING LEUKOREDUCED BLOOD SAMPLES
Described herein are methods and kits useful for the extraction and analysis of genomic DNA from leukoreduced blood or plasma samples.
1 . A method for genetic analysis, comprising: isolating DNA from a sample comprising leukoreduced blood; and analyzing the DNA for the presence of one or more genetic markers.
2 . (canceled)
3 . The method of claim 1 , wherein the step of isolating DNA comprises removing red blood cells from the sample prior to isolation of the DNA.
4 . The method of claim 1 , wherein the step of analyzing DNA comprises determining the presence or absence of one or more alleles of interest.
5 . The method of claim 4 , wherein the one or more alleles are polymorphisms in one or more human erythrocyte antigens.
6 . The method of claim 5 , wherein the antigens are selected from the group consisting of Rh, Kell, Duffy, Kidd, MNS, Lutheran, Dombrock, Cartwright, Diego, Colton, and Hemoglobin S.
7 . The method of claim 1 , wherein said leukoreduced blood contains less than 10 leukocytes/μl of sample.
8 . (canceled)
9 . The method of claim 1 , wherein said DNA is amplified prior to analysis.
10 . The method of claim 1 , wherein said DNA is isolated using magnetic beads.
11 . The method of claim 1 , wherein said DNA exhibits an A260/280 ratio of 1.5 or higher.
12 . The method of claim 1 , wherein said method isolates more than 50 ng DNA/ml of sample.
13 . The method of claim 1 , wherein said isolated DNA is subjected to multiplexed genotyping.
14 . The method of claim 1 , wherein the DNA is analyzed using an allele specific identification primer having a sequence specific region, a bead capture tag and a capture probe elongation template.
15 . The method of claim 14 , wherein the method comprises annealing a DNA to the allele specific identification primer.
16 . The method of claim 15 , wherein the step of annealing facilitates production of a single stranded DNA primed from the allele specific identification primer.
17 . The method of claim 16 , wherein said single-stranded DNA incorporates a labeled nucleotide.
18 . The method of claim 1 , wherein the DNA is analyzed using a plurality of capture beads.
19 . (canceled)
20 . (canceled)
21 . The method of claim 1 , wherein the step of isolation of DNA employs a kit selected from the group consisting of QIAamp Blood Mini Kit, QIAamp DNA Midi kit, QIAamp DSP Virus Spin Kit and QIAamp circulating nucleic acid Kit.
22 . The method of claim 1 , wherein the step of analyzing the DNA employs a kit selected from the group consisting of: HEA LR eMAP-S BeadChip Kit and HEA LR eMAP BeadChip Kit.
23 . A kit for analyzing extracted DNA comprising: an allele specific identification primer having a sequence specific region, a bead capture tag and a capture probe elongation template; a plurality of capture beads having bead capture probes attached thereto; and a container therefore.
24 . (canceled)
25 . A method of isolating DNA from a leukoreduced blood sample or plasma sample, comprising:
a. Providing a leukoreduced blood sample or plasma sample; and
b. Isolating more than 50 ng DNA/ml of sample.
26 - 34 . (canceled)