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Nucleic Acids Research logoLink to Nucleic Acids Research
. 2017 Dec 20;46(5):2697. doi: 10.1093/nar/gkx1296

ExoCET: exonuclease in vitro assembly combined with RecET recombination for highly efficient direct DNA cloning from complex genomes

Hailong Wang 1, Zhen Li 1, Ruonan Jia 1, Jia Yin 1, Aiying Li 1, Liqiu Xia 2, Yulong Yin 1,2,3, Rolf Müller 1,4, Jun Fu 1,, A Francis Stewart 5,, Youming Zhang 1,
PMCID: PMC5861451  PMID: 29272442

Nucleic Acids Research (2017) gkx1249, https://doi.org/10.1093/nar/gkx1249

The authors wish to make the corrections highlighted in bold to Table 1 of their article. The corrections have also been made in the published article and do not affect the results and overall conclusions of the work.

Table 1. Large genomic segments directly cloned from bacteria, mammalian cells and human blood with ExoCET.

Target Source Genome (Mb) Digestion enzymes Fragment (kb) Vector c.f.u. (/ml) Correct/checked
plu3535-3532 P. luminescens DSM15139 5.69 XbaI 38 pBAC2015 1815±132 12/12
plu2670 P. luminescens DSM15139 5.69 XbaI+XmaI 53 p15A 787±194 10/12
salinomycin cluster S. albus DSM41398 8.38 EcoRV 106 pBeloBAC11 425±91 2/24
salinomycin cluster S. albus DSM41398 8.38 Cas9 106 pBeloBAC11 260±14 1/24
Wnt4 Mouse melanoma B16 cell 2800.06 SwaI 45 p15A 76±16 8/25
Lmbr1l-Tuba1a Mouse melanoma B16 cell 2800.06 SwaI 53 p15A 52±6 1/12
Prkar1a Mouse melanoma B16 cell 2800.06 HpaI 8 p15A 205±17 10/12
IGFLR1-ARHGAP33 Human blood 3221.49 BstZ17I 41 p15A 275±76 5/48
ZBTB32-LIN37 Human blood 3221.49 NdeI 45 p15A 115±35 2/48
Dpy30 Mouse melanoma B16 cell 2800.06 BamHI+KpnI 8.7 p15A 273±18 9/12
DPY30 HEK 293T cell 3221.49 SpeI 9.1 p15A 40±10 17/24
DPY30 Human blood 3221.49 SpeI 9.1 p15A 45±2 5/24
Oct4-Venus Mouse R1 ES cells 2800.06 EcoRV+PacI 9.6 p15A 34±1 9/36
Nanog-Cherry Mouse R1 ES cells 2800.06 NdeI 13 p15A 49±12 17/54
Gata2-Venus Mouse GM8 ES cells 2800.06 BstZ17l 16.8 p15A 212±27 5/45
Mll4 (1) Mouse R1 ES cells 2800.06 SspI+SpeI 17.1 p15A 127±38 7+3/24
Mll4 (2) 323±65 2+2/36
Mll4 (3) 142±27 6+9/72
Mll4 (4) 483±91 3+5/36

All experiments were done in triplicate; c.f.u. includes standard deviation and fidelity was monitored by restriction analysis of the indicated number of colonies. For the Mll4 experiments, fidelity shows the targeted allele + wt allele/colonies examined. DNA analyses are shown in Supplementary Figure S6.


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