HydroLink gel electrophoresis: rapid electroblotting of dsDNA.
Blotting and probing of DNA, RNA and proteins after electrophoresis is a powerful technique for the study of the structure and function of biomolecules. Key factors in successful blotting experiments are efficiency of transfer, maintenance of the resolution obtained during gel electrophoresis, accuracy of the probes used and sensitivity of the detection method. We have recently developed a system for the high performance resolution of DNA with 10-fold greater capacity for sample loads than agarose or polyacrylamide. In the present study, we describe conditions for the rapid (less than one hour) and quantitative electrotransfer of DNA in the 100-23,000-base pair range, with subsequent conditions for probing of transfer membranes using radioactive or biotinylated probes. Our results suggest complete maintenance of the high-resolution characteristic of HydroLink gel electrophoresis and potentially increased sensitivity due to the high loading capacity in HydroLink gel electrophoresis.
Smith
C L
CL
AT Biochem, Malvern, PA 19355.
Ewing
C M
CM
Jain
T
T
Shorr
R G
RG
D001345:Autoradiography; D001710:Biotin; D004247:DNA / Q000032:analysis*; D015342:DNA Probes; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D004996:Ethidium; D005782:Gels; D006863:Hydrogen-Ion Concentration; D009691:Nucleic Acid Denaturation; D009757:Nylons; D010761:Phosphorus Radioisotopes; D011108:Polymers
D015342:DNA Probes; D005782:Gels; D009757:Nylons; D010761:Phosphorus Radioisotopes; D011108:Polymers; C061589:HydroLink; D001710:Biotin; D004247:DNA; D004996:Ethidium