I am hope to perform a PCR reaction on my targeted gene. Then do a gel electrophoresis and also purifying my DNA using Nucleospin Gel and PCR Clean- up kit. I am hoping to send my DNA for sequencing. My question is, after l have puriying my DNA what do l need to do to send it for sequencing ?. Will my DNA be fully ready for for sequencing after the purification step?
Thanks
Dear Fatna,
After purifying, do perform gel electrophoresis again just to make sure. If it's fine, then just proceed with sequencing.
I would not run the purified dna again if the original amplimer was clean . Just measure the amount of purified dna and check it is more than the company requests and send the dna and sequencing primers to the company.The company will have a fact sheet telling you everything you need to know about amounts and how to send the material and labelling etc
Dear Fatna,
After you purify the DNA, you can check the concentration and purity of the DNA using a NanoDrop machine. You should contact the company to know the volume they need for sequencing. Then send your DNA samples along with the sequencing primers. Because if you don't send the sequencing primers, then the company will charge for the primers as well. Please note that, when you send samples and primers, you should send it in a cool condition, so put a frozen ice-pack with the samples and put everything in a box.
Dear Fatna,
If you have NanoDrop machine in your lab then you can check the concentration and purity of DNA after the purification. But, If don't have then you can check on the gel and send for sequencing in a cool condition as per requirement of the sequencing company along with the primer.
As you asked, yes your DNA is fully ready for sequencing. Don't worry your result will be good.
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