How many fragments of DNA will result and what are their seq

How many fragments of DNA will result and what are their sequences, when using deoxythymidine triphosphate to sequence this fragment of gene? Give the primer sequence. Sequencing occurs on the indicated strand. 3\' AGTCACTGTTCGGCTAGTGCATGGTCG 5\' 5\' TCAGTGACAAGCCGATCACGTACCAGC 3\'

Solution

Answer:

DNA to be sequenced:

5\' TCAGTGACAAGCCGATCACGTACCAGC 3\'

The number of nucleotides in the above sequence is 27. So, number of fragments produced during sequencing will be 27. The sequences are given below:

G-5\'

CG-5\'

TCG-5\'

GTCG-5\'

GGTCG-5\'

TGGTCG-5\'

ATGGTCG-5\'

CATGGTCG-5\'

GCATGGTCG-5\'

TGCATGGTCG-5\'

GTGCATGGTCG-5\'

AGTGCATGGTCG-5\'

TAGTGCATGGTCG-5\'

CTAGTGCATGGTCG-5\'

GCTAGTGCATGGTCG-5\'

GGCTAGTGCATGGTCG-5\'

CGGCTAGTGCATGGTCG-5\'

TCGGCTAGTGCATGGTCG-5\'

TTCGGCTAGTGCATGGTCG-5\'

GTTCGGCTAGTGCATGGTCG-5\'

TGTTCGGCTAGTGCATGGTCG-5\'

CTGTTCGGCTAGTGCATGGTCG-5\'

ACTGTTCGGCTAGTGCATGGTCG-5\'

CACTGTTCGGCTAGTGCATGGTCG-5\'

TCACTGTTCGGCTAGTGCATGGTCG-5\'

GTCACTGTTCGGCTAGTGCATGGTCG-5\'

AGTCACTGTTCGGCTAGTGCATGGTCG-5\'

PCR primer having a universal sequencing primer binding site added to the 5´ end can be used as the primer for sequencing. Universal-tailed PCR primers are useful in the following scenarios:

(a) In conjunction with dye terminator chemistries (universal sequencing primers have good annealing characteristics)

(b) In conjunction with commercially available dye-labeled sequencing primers

The primer sequence should be located 50-60 bases upstream of the target DNA sequence and can be designed based on the upstream sequence.