RT Journal
A1 Andersson, B
A1 Ying, J H
A1 Lewis, D E
A1 Gibbs, R A
T1 Rapid characterization of HIV-1 sequence diversity using denaturing gradient gel electrophoresis and direct automated DNA sequencing of PCR products.
JF Genome Research 
JO Genome Research 
YR 1993 
FD May 01 
VO 2 
IS 4 
SP 293 
OP 300 
DO 10.1101/gr.2.4.293 
UL http://genome.cshlp.org/content/2/4/293.abstract 
AB A direct method for visualization and isolation of sequence variants of human immunodeficiency virus type 1 (HIV-1) utilizing denaturing gradient gel electrophoresis (DGGE) combined with automated direct DNA sequencing was developed. Two fragments from the env gene and one from the nef gene of HIV-1, which together constitute approximately 1.0 kb of sequence, were amplified by PCR and analyzed. HIV-1 variants from each region were resolved and excised from the gel; this was followed by direct sequencing of different viral variants. In 9 infected patients, a limited number of dominant sequence variants could be seen in the three regions, together with a faint background of minor variants. The use of DGGE makes it possible to obtain a direct estimate of overall HIV-1 sequence diversity within patient samples without an intermediate DNA cloning step.