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Molecular Biology help please! <3

Basically the lecture notes we have on this are pretty vague, and I'm really stuck :frown:

"You have 1.2kb of cDNA of a liver specific enzyme from rat. You wish to isolate the orthologous (similar sequence in a different species) coding sequence found in man. Explain the steps in this process.

Secondly, having isolated the human orthologue of the liver specific enzyme explain how you might then isolate the corresponding human genomic region."

Can anyone help?

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(edited 11 years ago)

Reply 1

alleycat393

You would probably need to use BLAST to get hold of the actual human cDNA and genomic sequences.

Use some kind of human liver cell line to isolate some RNA, make cDNA and use primers which span exon/exon boundaries in the human cDNA sequence to isolate and amplify the cDNA sequence.

For the genomic region, isolate DNA instead of RNA and use primers against the genomic sequence which span intron/exon boundaries to amplify the sequence.