Hi! I'm having problems with the tagment genomic DNA step of the Nextera mate-pair library preparation kit and I really don't know if the enzyme is working at all. I' attaching the bioanalyzer DNA 12000 results right after the tagment genomic DNA step. I tested two different lots of the enzyme but the results are the same, and I also don't think is a DNA quality problem because this DNA has been successfully used with the Nextera XT kit which also uses a tagmentation step.
I used 4 ug of DNA as input (good quality according to PFGE results with fragments longer than 50 kbp) and 4 uL (lane 1) or 8 uL (lane 2) of tagment enzyme from one lot and 4 uL from another lot (lane 3). Lane 4 was loaded with water.
There seems to be something in the 17000 bp region but I don't know if this long fragmented DNA or unfragmented DNA that migrated with the longer band of the ladder.
Can somebody please explain what happened with my fragmented DNA? Thanks!
I used 4 ug of DNA as input (good quality according to PFGE results with fragments longer than 50 kbp) and 4 uL (lane 1) or 8 uL (lane 2) of tagment enzyme from one lot and 4 uL from another lot (lane 3). Lane 4 was loaded with water.
There seems to be something in the 17000 bp region but I don't know if this long fragmented DNA or unfragmented DNA that migrated with the longer band of the ladder.
Can somebody please explain what happened with my fragmented DNA? Thanks!
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