In order to achieve this, robust surface for anchoring of DNA probes capable of withstanding elevated temperature for enzymatic reactions either in solid phase melting curve analysis or single dideoxynucleotide
incorporation will be developed.
After hybridization, the primer is extended by the polymerase with dideoxynucleotide
terminators in the absence of dNTPs.
Genomes were finished by using PCR and traditional dideoxynucleotide
sequencing (Sanger) to resolve gaps and repetitive and low coverage areas.
1,2) The Sanger method is also known as terminator sequencing because DNA fragments of varying lengths are synthesized by incorporating both nucleotides and dideoxyterminators (deoxyribonucleotide triphosphates [dNTPs] and dideoxynucleotide
triphosphates [ddNTPs], respectively).
DNA sequencing employs poisonous dideoxynucleotide
tri-phosphates (diNTPs) in a 1:1 ratio with the nonpoisonous dNTPs.
Sequencing was performed by fluorescent dideoxynucleotide
termination at the University of Utah core facility.
HIV RNA extraction, reverse transcription, amplification, dideoxynucleotide
sequencing, and algorithmic interpretation of the sequencing data are steps of:
Other approaches to improving sequencing technology involve methods that are independent of the Sanger dideoxynucleotide
chain termination reaction or of electrophoretic separation of the termination products.
5 [micro]l) for the sequence of interest, radiolabeled primers (1 [micro]l), dideoxynucleotide
triphosphates and DNA polymerase (9.
The amplified products were subjected to termination dideoxynucleotide
PCR products were sequenced by using the dideoxynucleotide
chain termination method with the ABI Prism BigDye Terminator Cycle Sequencing Reaction kit on an ABI Prism 3100 automated sequencer (Applied Biosystems, Naerum, Denmark).
Many molecular pathologists developed tests using traditional technologies, such as pyrosequencing, Sanger dideoxynucleotide
chain termination sequencing, or real-time polymerase chain reaction.