Method for determination of the length of the g-tail sequence and kit for the method

Abstract

A method of measuring the length of a G tail sequence, characterized by hybridizing the G tail of an nondenatured chromosomal DNA in a sample with a labeled DNA probe having a sequence complementary to the telomere repeat sequence, measuring chemiluminescence from the hybridized DNA probe, and determining the length of the G tail sequence from the measured value, and a kit used for use in such a method.

1 . A method of measuring the length of a G tail sequence, comprising hydrizing a G tail of an nondenatured chromosomal DNA in a sample with a labeled DNA probe having a sequence complementary to a telomere repeat sequence, measuring chemiluminescence from the hybridized DNA probe, and determining the length of the G tail sequence from the measured value. 
     
     
         2 . The method according to  claim 1 , wherein the sample is a cell pellet of blood, a cultured cell, a fresh tissue, a cryopreserved tissue or a formalin-fixed tissue. 
     
     
         3 . The method according to  claim 1  or  2 , wherein the fact that the chemiluminescence is based on the hybridization of the labeled DNA probe with the G tail sequence is confirmed by using an exonuclease. 
     
     
         4 . The method according to  claim 3 , wherein the exonuclease is an exonuclease I. 
     
     
         5 . The method according to any one of  claims 1  to  4 , wherein the label is an acridinium ester, luminol, isoluminol, pyrogallol, protohemin, aminobutylethyl-n-isoluminol or aminohexylethyl-n-ethyl-isoluminol. 
     
     
         6 . The method according to any one of  claims 1  to  5 , wherein the sequence complementary to the telomere repeat sequence is a base sequence represented by (CCCTAA) n  (n is an integer of 1 to 10). 
     
     
         7 . A kit for measuring the length of a G tail sequence, comprising at least a labeled DNA probe having a sequence complementary to an nondenatured telomere repeat sequence, a cytolytic solution, and a hydrolytic reagent. 
     
     
         8 . The kit according to  claim 7 , further comprising an exonuclease as a confirming agent. 
     
     
         9 . The kit according to  claim 7  or  8 , wherein the label is an acridinium ester, luminol, isoluminol, pyrogallol, protohemin, aminobutylethyl-n-isoluminol or aminohexylethyl-n-ethyl-isoluminol. 
     
     
         10 . The kit according to any one of  claims 7  to  9 , wherein the sequence complementary to the telomere repeat sequence is a base sequence represented by (CCCTAA) n  (n is an integer of 1 to 10). 
     
     
         11 . The method according to any one of  claims 1  to  7 , wherein the sample is a human- or mouse-derived sample.