Library prep with <100ng DNA?

[suludana]

Are you talking about the preparation of Genomic DNA?

[Efortier]

We routinely use 10 ng and 12 cycles without any problems. Just be careful to reduce the amount of adapters and PCR primers to take into consideration the lower amount of initial DNA.

[henry.wood]

I've fairly routinely used around 50ng. I've occasionally dipped down lower. According to my picogreen machine, my lowest is 5ng. We tweaked the method a bit, doing 18 cycles of PCR before the gel cut rather than 12 cycles afterwards. It doesn't always work, and sometimes we get a bit of PCR duplicatation, but 50ng is reasonably well behaved.

[sstarkenburg]

Thanks for the tip. Have you had any success/experience using Illumina's library prep kit at 100ng? If so, how many amplification cycles would you recommend?

[NextGenSeq]

Nextera's protocol only uses 50ng. However, the library insert size is not very tight with it. You could probably use almost any protocol if you increase the amplification cycle number. This would also increase artifacts tho.

[sstarkenburg]

What is the lowest amount of DNA I can get away with using for a library prep for a 36bp single read run on the IIX?