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Can I use more DNA than the amount indicated in the protocol?
Can I use more DNA than the amount indicated in the protocol?
Updated over a week ago

For the Ligation sequencing kit, the amount of sequencing adapter is fixed in the adapter ligation step and has been optimized for 0.2 pmol of end-prepped DNA.

If more DNA is taken into the step, the throughput will not increase accordingly and can waste precious input materials.

The Rapid sequencing kits have been optimized for an input of 400 ng of high molecular weight (HMW) DNA, i.e. >30 kb.

Since the transpose will tagment a finite number of times and there are more total DNA fragments in a larger amount, each fragment will be tagmented fewer times.

By using this mechanism, some users have increased the amount of HMW DNA used to produce longer read lengths at the expense of throughput.

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