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PCR Data Reading and Blotting Practice

1. PCR and RT-PCR

a. Transgenic banana plants
1. In-plant method
2. Protoplast cells that have been successfully transformed and those that have failed can be selected by
PCR technique
3. From the PCR test, the results were obtained in the form of several individuals who failed
to be transformed by seeing a positive sign on the PCR results screening. From the PCR
results of all lines it was known that the plants only contained the rcg3 or rcc2 genes
and did not contain the hpt gene and vice versa. Plants that are not transgenic plants
are plants that contain only one of the existing genes.
b. Tobacco plant
1. SGN-U213363 and SGN-U218605
2. SGN-U231627, SGN-U213276, SGN-U219218, SGN-U227655, and SGN-U223525
3. The plant variety that has the highest tolerance level is plant C then B and the
plant variety that has the lowest tolerance level is plant A. This can be seen from
the stability of the band as seen in the RT-PCR results
2. Southern Blot
1. Colonies 48, 49, 55, 58, 67, and 2
2. Colonies 3, 21, 25
3. Colony 21
3. Northern Blot
1. Spleen, P. Blood leukocyte, placenta, lung, skeletal muscle
2. Thymus, prostate, testis, colon (mucosal lining), brain, liver, kidney, pancrease
3. Pancreas
4. Pancreas
4. Western blot
1. Increased protein production occurs in COL6A2, vimentin, KIC9, ACTB, and HSP27.
Whereas in TYPH and KIC19 there was a decrease in protein.
2. Cancer growth has the possibility of being suppressed by proteins that have
increased after taking the drug, namely COL6A2, vimentin, KIC9, ACTB, and
HSP27.