Biomedical Engineering Reference
In-Depth Information
4. By alternating the antibiotic resistance gene between the hier-
archical levels of assembly (e.g., Ampicillin versus Kanamycin),
the cloning background resulting from undigested fi rst-level
plasmid DNA can be signifi cantly reduced.
Acknowledgement
The authors thank Veit Hornung for his support. This work is
fi nancially supported by the Bauer Fellows Program, SFB 670,
ETH Zurich, ERC starting grant, National Institutes of Health
(1R01CA155320-01), National Institute of General Medical
Sciences Grant for Centers of Systems Biology, and the German
National Academic Foundation.
References
1. Aslanidis C, Dejong PJ (1990) Ligation-
independent cloning of PCR products (LIC-
PCR). Nucleic Acids Res 18:6069-6074
2. Kodumal SJ, Patel KG, Reid R et al (2004)
Total synthesis of long DNA sequences: syn-
thesis of a contiguous 32-kb polyketide syn-
thase gene cluster. Proc Natl Acad Sci U S A
101:15573-15578
3. Reisinger SJ, Patel KG, Santi DV (2006) Total
synthesis of multi-kilobase DNA sequences
from
6. Geu-Flores F, Nour-Eldin HH, Nielsen MT et al
(2007) USER fusion: a rapid and effi cient method
for simultaneous fusion and cloning of multiple
PCR products. Nucleic Acids Res 35:e55
7. Gibson DG, Benders GA, Andrews-Pfannkoch
C et al (2008) Complete chemical synthesis,
assembly, and cloning of a
Mycoplasma genita-
lium
genome. Science 319:1215-1220
8. Gibson DG, Young L, Chuang RY et al (2009)
Enzymatic assembly of DNA molecules up to
several hundred kilobases. Nat Methods
6:343-345
9. Gibson DG, Benders GA, Axelrod KC et al
(2008) One-step assembly in yeast of 25 over-
lapping DNA fragments to form a complete
synthetic
Mycoplasma genitalium
genome.
Proc Natl Acad Sci U S A 105:20404-20409
10. Schmid-Burgk JL, Xie Z, Frank S et al (2012)
Rapid hierarchical assembly of medium-size
DNA cassettes. Nucleic Acids Res 40:e92
oligonucleotides.
Nat
Protoc
1:2596-2603
4. Aslanidis C, Dejong PJ, Schmitz G (1994)
Minimal length requirement of the single-
stranded tails for ligation-independent cloning
(LIC) of PCR products. PCR Methods Appl
4:172-177
5. Donahue WF, Turczyk BM, Jarrell KA (2002)
Rapid gene cloning using terminator primers
and modular vectors. Nucleic Acids Res 30:e95
