PREFINAL_DNA SEQUENCING

Question 1

• This refers to the order of the nucleotides in the DNA molecule

Answer 1

DNA Sequence

Question 2

Application of DNA sequencing in medical laboratory:

Answer 2

> Detection of mutation
Typing microorganism
Identifying human haplotypes
Designating polymorphism
Treatment strategies

Question 3

SEQUENCING METHOD:
- Direct determination of the order, or sequence of nucleotides in a DNA polymer.
- Most specific and direct method for identifying genetic lesions.

Answer 3

DIRECT SEQUENCING

Question 4

TYPES OF DIRECT SEQUENCING:

Answer 4

MANUAL & AUTOMATED FLUORESCENT SEQUENCING

Question 5

TYPES OF DIRECT SEQUENCING
- Allan M. Maxam & Walter Gilbert
- Requires a ds/ss version of the DNA region to be
sequenced with 1 end radioactively labeled (
32P)
- Sequencing proceeds in 4 separate reactions
- Template: labeled fragment

Answer 5

MANUAL SEQUENCING: CHEMICAL

Question 6

TYPE OF MANUAL SEQUENCING:
- Each Chain Termination (Sanger) Sequencing
- Frederick Sanger
- Uses dideoxynucleotides (ddNTPs) to determine
the order/sequence of nucleotides in a nucleic acid
 Primer complementary to DNA to be sequenced

Answer 6

MANUAL SEQUENCING: