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3. DNase-free filtered tips (10, 20, 200 and 1000 μL).


4. Electrophoresis gel tank system and agarose gel caster with
   well comb.


5. 1× Tris-acetate-EDTA (TAE) buffer.


6. 6× loading dye.


7. 100 bp DNA ladder.


8. Agarose powder.


9. Microcentrifuge tubes.


10. Polymerase chain reaction (PCR) tubes.


11. Primer set.


12. PCR mastermix (preferably high fidelity mastermix).


13. PCR hood.


14. 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