DNA sequencing

[desperate]

Hello everybody.

I am a medical doctor and my 'biology' experience is little. I am desperately seeking for help as I am running out of time.

Can anyone tell me how do I have to prepare genomic DNA extracted from human blood in order to have DNA-sequencing?
Until now, I have extracted the genomic DNA using mini spin columns and checked the extraction product with agarose gel electrophoreses.
What is the next step?
Should I estimate the DNA quantity extracted and how do I do this? Should I go on with PCR? If so, do I need branded primers or not? Do I have to estimate the PCR product quantity? etc?
I will not carry out the DNA sequencing process. A company will do it for me. I just have to prepare the samples.

Thanks in advance.

[aftabac]

Hi
you can directly seqence your DNA directly after extraction it from blood. there are two ways now you can go.
1-Do PCR on your specfic gene or part of DNA you want to sequence and send same primers for sequencing along with your PCR product.
2-amplify your gene of interest with PCR and then insert your gene into any of the cloning vector. now mostly people do Topo-cloning and these vector kit also provide you primer for sequencing. so you can send your vector with gene of interst along with primers to sequencing company.
for further detail i am going to send you link of a company Genewiz, you can check the quality of DNA and amount you required for sequencing as well as primer concentration on the site.
i do hope it will help you.

[Only registered users see links. ]

regards
aftab

[desperate]

Thank you very much for your kind assistance.

I think I will follow the first alternative as it is a process I already know.

Thanks again.