A. Safety operation in a PCR lab: 

• Hazardous chemicals.
• Safety rules for machines operation.
• Reagent preservation and lab security

• DNA structure.
• The function of DNA Polymerase.
• The principle of polymerase chain reaction (PCR).
• Basic requirements and design of PCR lab.
• Potential problems of PCR.

C. Introduction of PCR related reagents:

• DNA extraction kit
• IQ2000^TM WSSV Kit.
• Reagents for electrophoresis.

D. Introduction of PCR related equipment

E. Review of some major shrimp viral diseases:

• White spot syndrome virus (WSSV).
• Yellow-head virus/Gill-associated virus (YHV/GAV).
• Infectious hypodermal and haematopoietic necrosis virus (IHHNV).
• Taura syndrome virus (TSV).

• Avoid vertical transmission.
•  Avoid horizontal transmission.

A. Practice of DNA extraction

B. Practice of PCR operation procedure:

• Reaction condition.
• Reagents preparation.
• Reaction Procedure.

• Agarose gel preparation.
• Electrophoresis.
• Gel staining and data assay.

• Preservation of shrimp (tissue) samples.
• Preservation of DNA samples.
• Introduction of sample dissecting tools.
• Disinfection procedure of the sample dissecting tools.
• Sampling strategy for broodstocks.
• Sampling strategy for eggs and nauplii.
• Sampling strategy for PLs.
• Sampling strategy for juveniles in the grow-out pond.
• Sampling strategy for environmental factors.

A. Perform the assay by using real samples:

• To compare the sensitivity, reproducibility, and to practice more, it is necessary to go through the whole procedure by real samples again.
• Positive standards and negative control are required.
• Requirements of sample.

• Sampling, sensitivity, and statistical issue.
• False positive or false negative result.
• Reproducibility.
• Lab management.

• Environmental screening.
• Choosing the source of brooders and PLs.
•  Viral risk management for the grow-out pond.

A. Real samples operation:  to confirm the operation and process the other diagnostic systems.

B. Discussion.