Easy protocole for DNA sequencing - sequencing 1kb fragment (Apr/17/2007 )
can any one help me in sequencing my 1Kb pcr product. i dont know any thing about sequencing, and is my first time to do so, so please help me in finding an easy to use protocol for dna sequencing.
thank you all in advance
Lab. Gen. Che
What is the sample? PCR product? Plasmid? Concentration? Purity? Do you have a primer for the sequence? We need to know more.
its a pcr product pure and i have farword primer specific and revers primer degenarated.
Well, I'd recommend sending it out. Most sequencing centers want 10 ul of sample containing about 100 ng of DNA, mixed with about 10 pMol of primer. The sample should not contain too much salt of EDTA -- 10 mM Tris or TE is fine. Your sequencing center will tell you what they want precisely, and in what kind of tubes or plates.