Science Buddies: "Ask an Expert"

Hi,

I would like to know how to transfer a genomic DNA into a plasmid?

Thank You

Here is a couple websites that discuss experiments about transformation and plasmid.

http://humgen.wustl.edu/hdk_lab_manual/ ... mid08.html
http://www.genome.ou.edu/protocol_book/ ... _adxF.html

Michael

Hi juman,

One way to put a certain piece of DNA into a plasmid is to use restriction enzymes. Restriction enzymes are proteins that cut DNA at a particular place because they recognize certain base pair sequences.

Try looking up "restriction enzymes" online to see if that will help you get started with your project.

Feel free to post with more questions.

Cheers,
Heather

Hi,

The posts below are definitely accurately, but I think you need to carefully reword your question a bit further. I'd imagine that you are trying to stick just a gene or so into a plasmid, in which case the answers below are perfectly good. However, I think that the strict definition of genomic DNA implies the totality of the organism's genome, not just a single gene. In the case that you want to clone and/or sequence huge chunks of DNA in vectors, a better bet would be artificial chromosomes, as plasmids are very much size limited to around 10 kilobases (kb).

Bacterial artificial chromosomes are used for genomes of certain small-ish organisms (ex, certain large viruses) and hold about 100-300 kb, see more info here: http://en.wikipedia.org/wiki/Bacterial_ ... chromosome.

Yeast artificial chromosomes are for even bigger organisms and hold up to 1500 kb: http://en.wikipedia.org/wiki/Yeast_arti ... chromosome.

In addition, I'm sure that there are plenty more methods for even bigger genomes and sequences, so poke around on the web to find more.

Hope this helps!
Aaron Lin

it is a really hard question to answer. it becomes harder if you try to answer it in English(my first language is not English).
if you are really interested in these sort of topics i can recommend you Pro. Browns' book " Gene cloning and DNA Analysis"

best regard
kamran banan