2 replies to this topic

[ikwana]

its about my science project.

i have to search for allinase gene from garlic.

from what i understand, i have to extract the raw garlic and obtain its whole genome.

and then, from the whole genome, i do the pcr to amplify allinase gene.

but my lecturer said, "we can't isolates gene from whole genomic" why??? then how am i going to get the gene if it is not from whole genome?

my idea is like searching for a word from a dictionary.. is my concept wrong?

i only have plant mini kit( to obtain the whole genome from the garlic clove)and PCR kit (to amplify the allinase gene)
is there any other kit that i can use?

help me please... i'm confusedddddddd....

[Rsm]

Does this gene contain introns? Then you'll need to clone it from mRNA instead of DNA.

rsm

[claritylight]

Those 2 kits are all you need. Basically, you have the DNA of the garlic clove, which is the whole genome (introns + exons + other genomic elements), so your DNA contains the entire coding sequence for the garlic clove. You need to amplify this allinase gene (your target gene). Your PCR kit should contain the specific primers to amplify the allinase gene, so you don't need to isolate your gene from the whole genome. Your PCR kit (in a way) does the isolation for you because it will target specifically the allinase encoding gene by amplifying this region out of the whole genome. So when you run your PCR product on a gel, you should see that gene as a specific sized band, meaning you have PCR product of your allinase gene, your lecturer should tell you what size band to expect.

I'm not sure  why your lecturer was saying that you can't get your gene from the whole genome, because you can if your primers are specific enough for the gene

Hope this helps you.

ikwana, on 04 January 2011 - 01:01 AM, said:

its about my science project.

i have to search for allinase gene from garlic.

from what i understand, i have to extract the raw garlic and obtain its whole genome.

and then, from the whole genome, i do the pcr to amplify allinase gene.

but my lecturer said, "we can't isolates gene from whole genomic" why??? then how am i going to get the gene if it is not from whole genome?

my idea is like searching for a word from a dictionary.. is my concept wrong?

i only have plant mini kit( to obtain the whole genome from the garlic clove)and PCR kit (to amplify the allinase gene)
is there any other kit that i can use?

help me please... i'm confusedddddddd....

Edited by claritylight, 09 January 2011 - 08:31 PM.