2008
DOI: 10.1093/nar/gkn457
|View full text |Cite
|
Sign up to set email alerts
|

De novo DNA synthesis using single molecule PCR

Abstract: The throughput of DNA reading (sequencing) has dramatically increased recently due to the incorporation of in vitro clonal amplification. The throughput of DNA writing (synthesis) is trailing behind, with cloning and sequencing constituting the main bottleneck. To overcome this bottleneck, an in vitro alternative for in vivo DNA cloning must be integrated into DNA synthesis methods. Here we show how a new single molecule PCR (smPCR)-based procedure can be employed as a general substitute to in vivo cloning the… Show more

Help me understand this report

Search citation statements

Order By: Relevance

Paper Sections

Select...
3
1
1

Citation Types

2
19
0

Year Published

2009
2009
2020
2020

Publication Types

Select...
6
3

Relationship

2
7

Authors

Journals

citations
Cited by 27 publications
(21 citation statements)
references
References 21 publications
2
19
0
Order By: Relevance
“…For instance, candidate tissue-specific enhancers were found to be significantly enriched ( P -value ≤0.05, Fisher’s exact test) in binding sites for TFs within regulatory networks that are known to be important in the respective tissues, such as MYC and NFKB1 in heart, and HNF4A and SP1 in liver [46-49] (data not shown). The combined collection of enhancers predicted in the loci of the 200 most highly expressed genes in each of the tissues considered significantly overlap with ORegAnno, a manually curated collection of regulatory sequences [50], featuring a two-fold enrichment ( P -value <0.001, computed based on 1,000 randomized sequences genome-wide). Also, our enhancer predictions are enriched for specific epigenetic histone marks generally associated with distal transcriptional regulation, as suggested by 41% of predicted enhancers overlapping ChromHMM predictions for strong and weak enhancers (1.5-fold enrichment, P -value <0.001, computed based on 1,000 randomized sequences genome-wide [51]).…”
Section: Resultsmentioning
confidence: 99%
“…For instance, candidate tissue-specific enhancers were found to be significantly enriched ( P -value ≤0.05, Fisher’s exact test) in binding sites for TFs within regulatory networks that are known to be important in the respective tissues, such as MYC and NFKB1 in heart, and HNF4A and SP1 in liver [46-49] (data not shown). The combined collection of enhancers predicted in the loci of the 200 most highly expressed genes in each of the tissues considered significantly overlap with ORegAnno, a manually curated collection of regulatory sequences [50], featuring a two-fold enrichment ( P -value <0.001, computed based on 1,000 randomized sequences genome-wide). Also, our enhancer predictions are enriched for specific epigenetic histone marks generally associated with distal transcriptional regulation, as suggested by 41% of predicted enhancers overlapping ChromHMM predictions for strong and weak enhancers (1.5-fold enrichment, P -value <0.001, computed based on 1,000 randomized sequences genome-wide [51]).…”
Section: Resultsmentioning
confidence: 99%
“…Open-access proponents have implemented alternative databases. The ORegAnno database aims to collect a broad range of TFBS data, using a convenient data format that allows rapid submission [38]. The PAZAR database has a more complex data model, which allows rich annotation of the evidence underlying each TFBS, as well as a full description of the TFBS (conditions, interacting TFs, cell types, and so on) [39].…”
Section: Detection Of Cis-regulatory Elementsmentioning
confidence: 99%
“…However, since gene synthesis and cloning are preparative rather than analytical procedures, they (in contrast to NGS cell-free cloning) must also enable the physical retrieval of cloned DNA molecules in individual droplets. Cell-free cloning of synthetic genes was first accomplished by us ( 15 ) and later by others with higher throughput ( 16 18 ). However, it has not yet been accomplished using microfluidic technology.…”
Section: Introductionmentioning
confidence: 99%