Protocol Online logo
Top : New Forum Archives (2009-): : Tissue and Cell Culture

chk effect of cell line genes on normal dna - (May/18/2014 )

hi

i want to chk effect of cancerous cell line genome on a normal DNA so for that purpose wat shod i do

1stly how can i insert normal human dna in the cell line either directly or in form of serum or blood or clone

2ndly after how long i should xtract dna from cell line as i dont want to see protien.does  cell line contains all genes that are in human? if yes then is there any need of inserting normal human genes into it to chk  effect?

can any body guide me 

-madi_epi-

There are two methods you could use here - a form of somatic cell nuclear transfer where you remove the nucleus from a cell and insert a new one from a cancerous cell - time consuming, technically very very challenging but quite rigorous.

 

The other method would be to choose a few genes, clone them, and then transfect.  Note that any "normal" cell will have a copy of the gene(s) that you are trying to insert.  You may be able to delete them using something like Crispr/Cas or homologous recombination.

-bob1-