Fully automated nucleic acid extraction methods for tissue samples

Abstract

Automated methods for extracting nucleic acid from one or more tissue samples disposed on slides are disclosed. The methods utilize an automated slide staining apparatus that dispenses a plurality of nucleic acid extraction reagents onto the tissue sample, thus creating an extracted nucleic acid sample. The extracted nucleic acid sample may be used directly in downstream applications such as nucleic acid amplification or sequencing procedures, or may be further purified.

1 - 82 . (canceled) 
     
     
         83 . An automated method for isolating nucleic acid from one or more tissue samples, wherein the one or more tissue samples are disposed on one or more slides, wherein the method comprises the following steps:
 (a) applying a deparaffinization reagent onto the one or more tissue samples;   (b) applying an antigen retrieval buffer onto the one or more tissue samples;   (c) applying a protease buffer onto the one or more tissue samples; and   (d) heating the one or more samples, thereby generating an eluate, and   wherein the eluate comprises isolated nucleic acid from the one or more samples.   
     
     
         84 . The method of  claim 83 , further comprising the following step:
 (e) removing the eluate from the samples on the one or more slides.   
     
     
         85 . The method of  claim 84 , further comprising the following step:
 (f) purifying the isolated nucleic acid.   
     
     
         86 . The method of  claim 85 , further comprising the following step:
 (g) performing nucleic acid amplification on the isolated nucleic acid.   
     
     
         87 . The method of  claim 86 , further comprising the following step:
 (h) analyzing the isolated nucleic acid.   
     
     
         88 . The method of  claim 87 , wherein the analyzing comprises sequencing. 
     
     
         89 . The method of  claim 83 , wherein the nucleic acid is DNA. 
     
     
         90 . The method of  claim 83 , wherein the nucleic acid is RNA. 
     
     
         91 . The method of  claim 91 , further comprising the step of applying guanidine thiocynate onto the one or more tissue samples, after step (c). 
     
     
         92 . The method of  claim 83 , wherein the samples on the one or more slides are FFPE samples, fresh samples, or frozen samples. 
     
     
         93 . The method of  claim 83 , wherein the samples are microdissected samples. 
     
     
         94 . The method of  claim 83 , wherein the automated staining machine comprises one or more dispensers for dispensing the deparaffinization reagent, antigen retrieval buffer, and protease onto the samples on the one or more slides. 
     
     
         95 . The method of  claim 94 , wherein the automated staining machine further comprises a heating element for heating the samples on the one or more slides.