RT Journal Article
SR Electronic
T1 Low bias DNA for sustainable and efficient data storage
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.02.09.940411
DO 10.1101/2020.02.09.940411
A1 Yanmin Gao
A1 Xin Chen
A1 Jianye Hao
A1 Chengwei Zhang
A1 Hongyan Qiao
A1 Hao Qi
YR 2020
UL http://biorxiv.org/content/early/2020/02/10/2020.02.09.940411.abstract
AB In DNA data storage, the huge sequence complexity is challenging for repeatable and efficient information reading from massive DNA oligo pool. Here, we demonstrated that synthetic oligo pool comprising over ten thousand strands was largely skewed by PCR process due to its inherent mechanism of inefficient priming, product-as-template, error-spreading prone, which caused serious oligos dropout, over 50% oligos lost in repeated PCR amplification, much more fatal than base mutant for correct information retrieve. Therefore, we developed a new biochemical framework isothermal DNA reading (iDR) from large-scale oligo pool normalization (OPN) and isothermal amplification. Due to its priming-free and error-spreading proof, it achieved low-biased and stable amplification even for successive 10 times deep-reading without performance lost, the first repeatable DNA storage. Furthermore, the skewed oligo pool with uneven molecule copy number of each oligos was rectified by a total 1280 chemically synthesized OPN-probes, the largest scale oligo normalization so far, and the necessary amount of sequencing reads for perfect oligos retrieve was further largely decreased. These advanced features enable the iDR chemical framework being ideal for manipulating the huge sequence complexity and building the sustainable and efficient DNA storage “hardware”.