Bernhard Bein
bernhardbein.bsky.social
Bernhard Bein
@bernhardbein.bsky.social
PhD student in comparative genomics @ Senckenberg and Goethe University Frankfurt
Applying polymerase A/B and C read amplification to two mollusc and one arthropod sample, we saw that assembly contiguity was either higher or comparable with polymerase C reads, and assemblies were always more contigous when combining all libraries, probably due to complementary read drop-outs.
February 10, 2025 at 3:26 PM
Combining reads of polymerase A/B and C with HiC reads, we were able to assemble the maned sloth genome to chromosome level with a contig N50 of ~4.75 Mb, a 10x increase compared to polymerase A/B reads alone.

The assemblies 3.1 Gb size also surpasses the previous 500 Mb protocol size limit.
February 10, 2025 at 3:26 PM
We then used a different polymerase "C" to repeat amplification and sequencing, increasing contig N50 and compleasm/TOGA gene completness dramatically.

Aligning reads of B. torquatus to another sloth species, we saw obvious read drop-outs that polymerase C reads could cover:
February 10, 2025 at 3:26 PM
We therefore switched to the ultra-low input protocol, which contains a PCR amplification step of input DNA, employing two different polymerases we call A/B.

This way, we could sequence the maned sloth Bradypus torquatus to a coverage of 45X.

However, contig N50 of the assembly was below 1 Mb:
February 10, 2025 at 3:26 PM