Joining of long double-stranded RNA molecules through controlled overhangs.

by: NH Dekker, JA Abels, PT Veenhuizen, MM Bruinink, C Dekker

Nucleic Acids Res, Vol. 32, No. 18. (2004)

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Abstract

We describe two methods for creating long (>1 kb) dsRNA molecules with specific, user-controlled overhangs for efficient hybridization and ligation. The two methods create double-stranded RNA (dsRNA) molecules with 5' overhangs or with 3' overhangs using T7 RNA polymerase (T7 RNAP) in transcription reactions of carefully designed PCR products. Primers utilized in the PCR reactions provide the template for the desired dsRNA overhangs. These methods provide complete control of the length and the sequence of the overhangs. This supplies a tool which is particularly lacking in dsRNA biochemistry given the absence of restriction endonucleases active on these substrates.

@article{citeulike:1828134, abstract = {We describe two methods for creating long (>1 kb) dsRNA molecules with specific, user-controlled overhangs for efficient hybridization and ligation. The two methods create double-stranded RNA (dsRNA) molecules with 5' overhangs or with 3' overhangs using T7 RNA polymerase (T7 RNAP) in transcription reactions of carefully designed PCR products. Primers utilized in the PCR reactions provide the template for the desired dsRNA overhangs. These methods provide complete control of the length and the sequence of the overhangs. This supplies a tool which is particularly lacking in dsRNA biochemistry given the absence of restriction endonucleases active on these substrates.}, address = {Kavli Institute of Nanoscience, Faculty of Applied Sciences, Delft University of Technology, Lorentzweg 1, 2628 CJ Delft, The Netherlands. Nynke.Dekker@mb.tn.tudelft.nl}, author = {Dekker, N. H. and Abels, J. A. and Veenhuizen, P. T. and Bruinink, M. M. and Dekker, C. }, citeulike-article-id = {1828134}, issn = {1362-4962}, journal = {Nucleic Acids Res}, number = {18}, posted-at = {2007-10-27 07:43:08}, priority = {2}, title = {Joining of long double-stranded RNA molecules through controlled overhangs.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/15475386}, volume = {32}, year = {2004} }

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TY - JOUR ID - citeulike:1828134 L3 - citeulike-article-id:1828134 TI - Joining of long double-stranded RNA molecules through controlled overhangs. JF - Nucleic Acids Res VL - 32 IS - 18 AD - Kavli Institute of Nanoscience, Faculty of Applied Sciences, Delft University of Technology, Lorentzweg 1, 2628 CJ Delft, The Netherlands. Nynke.Dekker@mb.tn.tudelft.nl SN - 1362-4962 N2 - We describe two methods for creating long (>1 kb) dsRNA molecules with specific, user-controlled overhangs for efficient hybridization and ligation. The two methods create double-stranded RNA (dsRNA) molecules with 5' overhangs or with 3' overhangs using T7 RNA polymerase (T7 RNAP) in transcription reactions of carefully designed PCR products. Primers utilized in the PCR reactions provide the template for the desired dsRNA overhangs. These methods provide complete control of the length and the sequence of the overhangs. This supplies a tool which is particularly lacking in dsRNA biochemistry given the absence of restriction endonucleases active on these substrates. AU - Dekker, NH AU - Abels, JA AU - Veenhuizen, PT AU - Bruinink, MM AU - Dekker, C PY - 2004/// UR - http://view.ncbi.nlm.nih.gov/pubmed/15475386 ER -

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