That`s half right. Is not as easy and fast , as one can guess.
Sorry if there is any mistake, but I think that you can get an idea of the whole process. Also, take in mind that to perform this process is not as easy as it may seem (specially for the gel part)
El viernes, 4 de noviembre de 2016, 9:15:15 (UTC+1), Hugues escribió:
-- The trick is that you need to label the last nucleotide of your "fragment" sequence in order to know "which nucleotide goes next".
In short, you cut your DNA in random length fragments (10, 100, 56, 2, 17bp, etc..). After this, you label each fragment at the end extreme (isotope, fluorescence, whatever). Then, you run the electrophoresis to separate the different fragments lengths. After this, you start reading in your gel nucleotide by nucleotide, so if you have a 1000bp fragment... well, enjoy the reading ;)
Sorry if there is any mistake, but I think that you can get an idea of the whole process. Also, take in mind that to perform this process is not as easy as it may seem (specially for the gel part)
About the miniPCR kit. I have one of those PCR and it works like a professional one (a side that it makes a LOT of noise due to the fan cooling system). So, yes: in theory you could used to sequence (in case you know and can label your fragments). BUT, to run the gels you should use an acrilamide gel, not agarose (which means you need "another" electrophoresis system).
My recommendation: purify your samples and send it to any company (ex: BGI, Macrogen...) or contact to any university's core facility.
It will be cheaper, faster and will save you from a lot of headaches. And, if there is any error, you will know and (most probably) will get a re-sequencing for free.
El viernes, 4 de noviembre de 2016, 9:15:15 (UTC+1), Hugues escribió:
But wait, i'm confused now. I thought we could read a DNA sequence with gel electrophoresis, no ?Isn't what we can Sanger sequencing, with the help of dideoxynucleotides ?example here:but i'm new to this, so maybe i missed something ? or this method works but it's too long and nobody uses it anymore ?
On Friday, 4 November 2016 04:07:59 UTC+1, SC wrote:That isn't a sequencer. It's a PCR machine and an electrophoresis unit. You won't be able to get sequence out of it.
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