Development of a modified method for sequencing high G:C regions in chicken anemia virus genome.
Two areas in the chicken anemia virus (CAV) genome have high G:C content with secondary structures. These two G:C rich areas could not be sequenced with Perkin Elmer's Big Dye Terminator Cycle Sequencing Kit. Several modifications were carried out to solve the problem. Finally, a package of modified method was developed to sequence the high G:C areas. The result showed that the Perkin Elmer's Big Dye Terminator Cycle Sequencing Kit with the normal procedures are not suitable for sequencing the high G:C regions of the CAV genome. The present developed method made the Perkin Elmer's Kit useful for the first time to sequence the G:C rich hairpin structures of the CAV genome. The system may be useful to sequence all other G:C rich DNA templates.
Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
S M Z H
SM
Chowdhury
A R
AR
Omar
I
I
Aini
M
M
Hair-Bejo
A A
AA
Jamaluddin
K L
KL
Wan
Y
Y
Kono
D000818:Animals; D001483:Base Sequence; D017727:Chicken anemia virus / Q000235:genetics*; D003001:Cloning, Molecular; D003596:Cytosine; D005821:Genetic Techniques; D016679:Genome, Viral; D006147:Guanine; D008957:Models, Genetic; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA / Q000379:methods*; D014407:Tumor Cells, Cultured
D006147:Guanine; D003596:Cytosine