title: [Gel immobilization of human genome].
abstract: OBJECTIVE
  To develop a solid phase PCR method by covalent single point immobilization for recycle utilization of human genome.
  
  
  METHODS
  Polymethacrylamide gel was selected as a solid PCR carrier based on DNA-hydrogel copolymer chemistry presented by Mirzabekov. (CH2)6NH2 amino-modified PCR product and randomly fractured formic acid-modified plasmid pGEM-T-HLA-G were used as templates. The specificity of the attachment chemistry was characterized by acrylamide gel electrophoresis, and the thermal stability of method was demonstrated by PCR. This method was applied for the recycle utilization of human genome. Sequencing was used to exclude the possibility of introduced mutations during modification and immobilization procedures.
  
  
  RESULTS
  The PCR detections of plasmid DNA and human genome DNA immobilized by polymethacrylamide gel was successful. The thermal stability of method was successfully demonstrated by performing PCR after 16 rounds of standard 36 PCR cycles. And the sequencing was found no mutation.
  
  
  CONCLUSIONS
  The DNA immobilization method with polymethacrylamide gel as a solid phase carrier is stable and specific, which can be a possible approach for realizing recycle utilization of human genome for whole-genome sequencing and SNP detection.
authors.affiliation: Taizhou Hospital Affiliated to Wenzhou Medical College, Linhai, China.
authors.forename: Yingqiu
authors.initials: Y
authors.lastname: Pan
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Shuqing
authors.initials: S
authors.lastname: Chen
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D015894:Genome, Human; D006801:Humans; D020100:Hydrogels; D061925:Immobilized Nucleic Acids / Q000032:analysis*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D020100:Hydrogels; D061925:Immobilized Nucleic Acids