Systems and Methods for Whole Genome Amplification

Abstract

Provided herein are systems and methods for whole genome amplification and sequencing. In particular, provided herein are systems and methods for detection of nucleic acid variants (e.g., rare variants) in limited samples.

1 .- 13 . (canceled) 
     
     
         14 . A system, comprising:
 a) a phi29 polymerase;   b) a heat stable polymerase; and   c) at least one nucleic acid primer, wherein each of said primers has the same   
       or different nucleic acid sequence. 
     
     
         15 . The system of  claim 14 , wherein said system further comprises dNTPs and a buffering agent. 
     
     
         16 . The system of  claim 14 , further comprising reagent for performing a nucleic acid sequence assay. 
     
     
         17 . The system of  claim 14 , wherein at least a subset of said primers comprises a sample barcode, molecular barcode, label, or tag sequence. 
     
     
         18 . The system of  claim 14 , wherein said system further comprises a microfluidic chip comprising a plurality of wells. 
     
     
         19 . The system of  claim 14 , wherein at least one of said at least one primer comprises 3′ ends comprising 3-8 nucleotides designed to bind to intron/exon boundaries in a target nucleic acid. 
     
     
         20 . The system of  claim 19 , wherein said nucleotides are selected from the group consisting of YAG, YAC, RTC and RTG. 
     
     
         21 . The system of  claim 14 , wherein at least one of said at least one primer comprises 3′ ends comprising 3-8 nucleotides that bind to repeat element consensus sequences.