219
- DNase-free filtered tips (10, 20, 200 and 1000 μL).
- Electrophoresis gel tank system and agarose gel caster with
well comb. - 1× Tris-acetate-EDTA (TAE) buffer.
- 6× loading dye.
- 100 bp DNA ladder.
- Agarose powder.
- Microcentrifuge tubes.
- Polymerase chain reaction (PCR) tubes.
- Primer set.
- PCR mastermix (preferably high fidelity mastermix).
- PCR hood.
- 96-Well PCR plate.
Table 3
Current main purposes of Sanger sequencing
Applications Example Ref.
Human leukocyte antigen (HLA)
typing
Speeding up the process of identifying a match in
unrelated donors for hematopoietic stem cell
transplantation
[ 23 ]
Multiple region sequencing Detection of HtrA Serine Peptidase 2 (HTRA2) and
anoctamin 3 (ANO3) mutation in patients with
essential tremor
[ 24 ]
Next-generation sequencing (NGS)
result validation
Alternate-method confirmation of mutations found in
patients with osteogenesis imperfecta
[ 25 ]
Confirmation of potential causative variants for Leber
congenital amaurosis
[ 26 ]
Plasmid sequences, inserts and/or
mutations confirmation
Deletion of dystrophin was confirmed with Sanger
sequencing in Duchenne muscular dystrophy
patients
[ 27 ]
Single disease-causing genetic variant
identification
Detection of breast cancer type 2 susceptibility
protein (BRCA2) mutation (c.1313delAAGA) in
the Moroccan population as first-line screening for
patients with breast and ovarian cancer
[ 28 ]
Targeted genomic regions with large
sample size
Detection of multidrug- and extensively drug
resistant tuberculosis in sputum specimen
[ 29 ]
Germline testing of BRCA1/2 in blood samples of
patients with high-grade serous ovarian cancer
[ 30 ]
Detection of telomerase reverse transcriptase (TERT)
mutation in patients with thyroid carcinoma
[ 31 ]
Single Gene Mutation in Esophageal Adenocarcinoma