Method of Measuring Telomere Length
Disclosed herein is a novel method of measuring telomere length comprising determining the DNA content (Dx) of a sample, determining the telomeric content (T) of a sample, and determining the value of T/Dx.
1 . A method of measuring telomere length comprising the steps of:
(a) determining the DNA content (Dx) of a sample;
(b) determining the telomeric content (T) of a sample; and
(c) determining the value of T/Dx.
2 . The method of claim 1 , wherein the sample has been applied to a nylon membrane.
3 . The method of claim 1 , wherein the determination of Dx is performed by applying to the sample a composition capable of visualizing the total nucleic acid in the sample.
4 . The method of claim 3 , wherein the composition is a DNA blot stain.
5 . The method of claim 3 , wherein the composition is SYBR DX DNA blot stain.
6 . The method of claim 1 , wherein the determination of T is performed by hybridizing the sample with a labeled telomeric probe.
7 . The method of claim 6 , wherein the labeled telomeric probe comprises the DNA sequence CCCTAA.
8 . The method of claim 6 , wherein the probe is labeled with digoxigenin.
9 . The method of claim 1 , wherein the sample comprises DNA isolated from leukocytes.
10 . A kit for measuring telomere length comprising:
(a) DNA blot stain;
(b) labeled telomere probe; and
(c) one or more membranes.
11 . The kit of claim 10 , further comprising at least one buffer.
12 . The kit of claim 10 , further comprising instructions for utilizing the kit for practicing the method disclosed herein.
13 . The kit of claim 10 , further comprising a composition capable of detecting the labeled telomere probe.
14 . The kit of claim 10 , further comprising CDP-Star solution, DNase-free water, and standard DNA.
15 . A kit for measuring telomere length comprising:
(a) SYBR DX DNA blot stain;
(b) DIG-labeled telomere probe;
(c) anti-DIG-alkaline phosphatase antibody conjugate;
(d) one or more nylon membranes.
(e) denaturing buffer, neutralizing buffer, saline-sodium citrate buffer (SSC), tris-borate-EDTA buffer (TBE), hybridization buffer, washing buffer, maleic acid buffer, blocking buffer, and detection buffer;
(f) CDP-Star solution;
(g) DNase-free water; and
(h) standard DNA.