Esophageal Adenocarcinoma Methods and Protocols

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

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

  3. 1× Tris-acetate-EDTA (TAE) buffer.

  4. 6× loading dye.

  5. 100 bp DNA ladder.

  6. Agarose powder.

  7. Microcentrifuge tubes.

  8. Polymerase chain reaction (PCR) tubes.

  9. Primer set.

  10. PCR mastermix (preferably high fidelity mastermix).

  11. PCR hood.

  12. 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
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