Esophageal Adenocarcinoma Methods and Protocols

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1. Optimize the annealing temperature of the primer sets designed
   by running a conventional PCR with gradient temperature.


2. PCR hood should be exposed to ultraviolet lights for 25 min
   prior to using it.


3. Prepare all PCR materials within the PCR hood to reduce
   chances of contamination.


4. Dissolve 5 g of agarose powder in 100 mL of 1× TAE buffer
   inside a 250 mL conical flask (see Note 7).


5. Put the mixture in the microwave oven to heat for 2 min (see
   Note 8).

3.3 Annealing
Temperature
Optimization
for ddPCR Primers
on Conventional PCR

Fig. 1 Outlook of Primer-BLAST Website for design of primers

DNA Copy-Number in Esophageal Adenocarcinoma