Criteria for Successful SMGT Technique:

• Quality of semen sample
• DNA uptake rate which depends on viability and motility of sperm cells.

Limitation:

• Sperm cells take up any DNA from the environment which reduces the specificity of the vector
• High amount of foreign DNA inhibits sperm movement.
• Large amount of DNA may get fragmented during the activation of sperm nucleus.

 
6-3.4 Characterization of Transgenic Animals:
After successful introduction of transgene into the host animal, the resulting transgenic animal should be kept under observation to study the stability and expression pattern of transgene inside host system. Continued expression of the product depends on two aspects of stability of the transgene:

• The structural stability or integrity of the transgene.
• The stability of transgene expression.

A. Genetic Stability:
In presence of multiple integration sites in the host genome, there may be occurrence of rearrangement or deletion of integrating transgene as well as host genome.To overcome this, stability of the transgene should be regularly monitored by tests like Southern blots, sequencing etc

B. Stability of expression:
The stability of expression of a transgene product can vary depending on interaction of the genetic background of the host animals or imprinting effects due to maternal or paternal inheritance. Stability of expression of the transgene product should be monitored over the productive life of the transgenic animal. Methods may include Northern blots, RT-PCR, DNase protection assay or other appropriate techniques.