Hello,
I'm trying to use an old Nextera kit from before the company was acquired by Illumina. I have tried using the kit's HMW buffer on 15 ng of the included lambda control DNA.
After tagmentation and limited-cycle PCR, I get the attached size distribution. This is not anything like the size distribution I expected from the rather terse instruction manual, nor is it similar to the ones pictured here:
Would the correct interpretation be that tagmentation failed or worked only poorly, or am I looking at something else entirely here? Besides bioanalyzer traces, what are other standard controls I might do to validate my library? I've thought about trying to PCR up random fragments, but it seems difficult. I'll appreciate any input at all.
Carl Wivagg
I'm trying to use an old Nextera kit from before the company was acquired by Illumina. I have tried using the kit's HMW buffer on 15 ng of the included lambda control DNA.
After tagmentation and limited-cycle PCR, I get the attached size distribution. This is not anything like the size distribution I expected from the rather terse instruction manual, nor is it similar to the ones pictured here:
Would the correct interpretation be that tagmentation failed or worked only poorly, or am I looking at something else entirely here? Besides bioanalyzer traces, what are other standard controls I might do to validate my library? I've thought about trying to PCR up random fragments, but it seems difficult. I'll appreciate any input at all.
Carl Wivagg
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