| Chapter title |
SLIC: A Method for Sequence- and Ligation-Independent Cloning.
|
|---|---|
| Chapter number | 5 |
| Book title |
Gene Synthesis
|
| Published in |
Methods in molecular biology, February 2012
|
| DOI | 10.1007/978-1-61779-564-0_5 |
| Pubmed ID | |
| Book ISBNs |
978-1-61779-563-3, 978-1-61779-564-0
|
| Authors |
Li MZ, Elledge SJ, Mamie Z. Li, Stephen J. Elledge, Li, Mamie Z., Elledge, Stephen J. |
| Abstract |
We describe here a method for sequence- and ligation-independent cloning (SLIC). SLIC uses an exonuclease, T4 DNA polymerase, to generate single-stranded DNA overhangs in insert and vector sequences. These fragments are then assembled in vitro and transformed into Escherichia coli to generate recombinant DNA of interest. SLIC inserts can also be generated by incomplete PCR (iPCR) or mixed PCR. As many as five inserts can be assembled in one reaction simultaneously with great efficiency using SLIC. SLIC circumvents sequence constraints for recombinant DNA using standard restriction enzyme-mediated cloning and previous ligation-independent cloning methods and provides a new approach for the efficient generation of recombinant DNA. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 2 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 2 | 100% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 6 | 2% |
| Germany | 2 | <1% |
| United Kingdom | 2 | <1% |
| Ireland | 1 | <1% |
| Australia | 1 | <1% |
| Canada | 1 | <1% |
| Switzerland | 1 | <1% |
| Denmark | 1 | <1% |
| New Zealand | 1 | <1% |
| Other | 0 | 0% |
| Unknown | 360 | 96% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 84 | 22% |
| Researcher | 67 | 18% |
| Student > Bachelor | 58 | 15% |
| Student > Master | 48 | 13% |
| Student > Doctoral Student | 14 | 4% |
| Other | 37 | 10% |
| Unknown | 68 | 18% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 127 | 34% |
| Biochemistry, Genetics and Molecular Biology | 120 | 32% |
| Chemistry | 15 | 4% |
| Immunology and Microbiology | 8 | 2% |
| Medicine and Dentistry | 7 | 2% |
| Other | 19 | 5% |
| Unknown | 80 | 21% |