PrbbBB:Oligo Annealing: Difference between revisions
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(New page: Back to all protocols ==Overview== Short to medium length (up to 120 bp) fragments of dsDNA can be generated directly from two complementary single-stranded oligonu...) |
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* Silver lab protocol (using water bath): [[Silver:_Oligonucleotide_Inserts]] | * Silver lab protocol (using water bath): [[Silver:_Oligonucleotide_Inserts]] | ||
* brief mentioning in Li & Elledge SLIC paper <cite>Li2007</cite> | * brief mentioning in Li & Elledge SLIC paper <cite>Li2007</cite> | ||
* [http://www.protocol-online.org/cgi-bin/prot/view_cache.cgi?ID=1282 Web protocol] | |||
The idea is to heat up the mixture to close to 100 C and let it slowly cool down to room temperature over the course of one hour. | The idea is to heat up the mixture to close to 100 C and let it slowly cool down to room temperature over the course of one hour. | ||
Revision as of 03:54, 21 July 2010
Overview
Short to medium length (up to 120 bp) fragments of dsDNA can be generated directly from two complementary single-stranded oligonucleotides. Custom overhangs for ligation or In-Fusion can be introduced as well, simply by having some non-complementary ends. The two single-stranded Oligos then need to be annealed into a double-stranded fragment.
See also:
- Silver lab protocol (using water bath): Silver:_Oligonucleotide_Inserts
- brief mentioning in Li & Elledge SLIC paper [1]
- Web protocol
The idea is to heat up the mixture to close to 100 C and let it slowly cool down to room temperature over the course of one hour.
Materials
- Thermocycler
- some buffer
Procedure
- step 1
- step 2
Notes
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
raik: We are just starting to use this protocol -- share your experience!
References
- Li MZ and Elledge SJ. Harnessing homologous recombination in vitro to generate recombinant DNA via SLIC. Nat Methods. 2007 Mar;4(3):251-6. DOI:10.1038/nmeth1010 |
Contact
or instead, discuss this protocol.
