Discussion Category: DNA Sequence Analyzers
regarding whole exome sequencing
Im doing whole exome sequencing on Ion torrent platform. I started with 1.4microgram/microliter of stock DNA. The fragmentation of the DNA was done correctly as i can see the smear on E-gel near 130-150basepair. Next i proceed with the adapter ligation of the library and then to the size selection step of the library preparation. On library preparation step i picked up the library of 210 basepair size. Then i proceed with the size selected library amplification. When i checked the Qubit concentration of my amplified library, it was below 4 ng/microliter. This concentration must be near 10 or 11 ng/microlitre. Can anyone tell me where is the flaw in this whole experiment.
Reply bySometimes the accurancy of instrument can affect the result of your experiment, if you can not find flaw in your process, you can check if something goes wrong with the instrument in you exome sequenncing experiment.