FISH Training Carried Out at NIAS-V on 3 – 23 September 2018

Establishing reproduction/breeding techniques for PERV-free or low copy-PERV Vietnamese native pigs is one of the Project activities. FISH, or Fluorescence In Situ Hybridization is one of the methods used in the Project to detect and localize the presence of PERV. The Project mainly uses Real Time-PCR for DNA analysis, but FISH is also recommended as a complementary method.

FISH training was first carried out at NIAS-V in July 2017 and this second training was organized as a follow-up. During this training, FISH was performed for PERV at interphase and metaphase, then checked signals using fluorescence microscope equipped by the Project in NIAS-V.

During the training, the trainees performed metaphase spread using the protocol. Attached are the photos acquired of metaphase spread during the training.

Metaphase spread

FISH was then performed using DNA samples of low copy-PERV pigs and commercial pig as a comparison. Attached are the photos of FISH for PERV in 4-cell embryos at interphase in the commercial pig.

 

 

A488 (probe for PERV stain)

 

 

 

 

DAPI (nuclear stain)

Merge

 

 

 

 

 

 

After the training course, the trainees could perform FISH using cells at interphase and metaphase, and check the samples using fluorescence microscope equipped by the Project in NIAS-V on their own.

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