Transgenomic

SURVEYOR Mutation Detection Kits and the SURVEYOR Check-It Kit provide reagents and protocols for the detection of mutations in DNA. All of these kits are based on the use of SURVEYOR Nuclease, a mismatch-specific endonuclease derived from celery, that has been shown to recognize and cleave all types of mismatches arising from the presence of single nucleotide polymorphisms (SNPs) or from small insertions or deletions. The protocol is based on the generation of PCR products that are subsequently hybridized to generate mismatches in heteroduplexed DNA which is then treated and cleaved by SURVEYOR Nuclease as outlined in the four-step protocol below.

Find any mutation even in longer fragments ranging up to 3.5 kb. SURVEYOR Nuclease fragments of PCR products derived from any genomic or cDNA sequences can be analyzed employing a variety of analytical platforms, and pooling can be used to increase throughput.

SURVEYOR Mutation Detection Kits

Designed to detect mutations in DNA derived from a variety of organisms including bacteria, fungi, plants and animals
Kits are provided for analysis by
- WAVE and WAVE HS Systems
- Standard Gel Electrophoresis
- LI-COR 3200 and 4200 Instruments
- Fluorescent Capillary Electrophoresis
PCR products can be prepared using high fidelity thermostable DNApolymerases such as Optimase Polymerase. Such amplicons give rise to the least amount of background fragments and permit greater depth of pooling. For standard assays, T-Taq DNA Polymerase can also be used.
For Standard Gel Electrophoresis the sensitivity is also enhanced by the use of Transgenomic's TransOneK or TransFiveK Agarose. The exceptionally low background fluorescence of these agaroses permits the detection of mutations that may be present at a low percentage such as 3%

SURVEYOR Check-It Kit for Clone Sequence Validation

Designed to detect mutations in plasmid DNA derived from bacterial colonies using colony PCR techniques
A Taq polymerase such as Transgenomic's T-Taq DNA Polymerase is needed for the colony PCR amplification step.
SURVEYOR Nuclease cleavage products are analyzed by standard gel electrophoresis.

Features	Benefits
SURVEYOR Nuclease provided as a standardized enzyme formulation	Detection of any base substitution and small insertion or deletion Fragments that are longer than those that can be sequenced in one run can be scanned for mutations in a single reaction Pooling of samples allows the detection of 1 mutant allele out of 32 and in some cases can increase the analysis throughput Versatility of the technique facilitates the use of various analytical platforms
SURVEYOR Nuclease provided in a kit format	SURVEYOR Nuclease is provided as a standardized quality-controlled enzyme formulation in a ready-to-use kit that can be used without further batch-to-batch optimization.
Reaction buffer, positive control plasmids and PCR primers and water included	Reaction buffers and positive controls are part of the kit and allow the user to set up and check the assays to obtain reproducible results.
User Guide with detailed instructions included	User Guides provide detailed instructions and troubleshooting guides Technical support is always available
SURVEYOR Nuclease available in bulk	Larger quality controlled amounts of SURVEYOR Nuclease are available in a bulk format upon request.

Catalog No.	Order	Product	Notes
706025	Order	SURVEYOR Mutation Detection Kit - S25	For Standard Gel Electrophoresis - 25 reactions
706020	Order	SURVEYOR Mutation Detection Kit - S100	For Standard Gel Electrophoresis - 100 reactions
706015	Order	SURVEYOR Mutation Detection Kit - F25	For Fluorescent Capillary Electrophoresis - 25 reactions
706010	Order	SURVEYOR Mutation Detection Kit - F100	For Fluorescent Capillary Electrophoresis - 100 reactions
706035	Order	SURVEYOR Mutation Detection Kit - W25	For WAVE and WAVE HS Systems - 25 reactions
706030	Order	SURVEYOR Mutation Detection Kit - W100	For WAVE and WAVE HS Systems - 100 reactions
706005	Order	SURVEYOR Mutation Detection Kit - L25	For LI-COR 4200 and 4300 Instruments - 25 reactions
706000	Order	SURVEYOR Mutation Detection Kit - L100	For LI-COR 4200 and 4300 Instruments - 100 reactions
706040	Order	SURVEYOR Check-It Kit	For Clone Sequence Validation - 100 reactions

LI-COR is a trademark of LI-COR, Inc.

Shipping
SURVEYOR Nulcease Kits are shipped on dry ice. Please store them at -15 to -30 degrees C. Minimum shelf life expected is 6 months after delivery if stored as directed.

Quality Control
All SURVEYOR Nuclease Kits are manufactured under ISO 9001 compliance. Quality control of all individual components and the complete assay kit is performed.

References
References identified with ** can also be found in our database of journal articles. They discuss Transgenomic products and technology.

Caldwell, D.G., McCallum, N., Shaw, P., Muehlbauer, G.J., Marshall, D.F. and Waugh, R. (2004) A structured mutant population for forward and reverse genetics in Barley (Hordeum vulgare L.). Plant J. 40, 143-150. Abstract
Colbert, T. Till, B.J., Tompa, R., Reynolds, S., Steine, M.N., Yeung, A.T., McCallum, C.M., Comai, L. and Henikoff, S. (2001) High-throughput screening for induced point mutations. Plant Physiol. 126, 480-484. Abstract
Greene, E.A., Codomo, C.A., Taylor, N.E., Henikoff, J.G., Till, B.J., Reynolds, S.H., Enns, L.C., Burtner, C. Johnson, J.E., Odden, A.R., Comai, L. and Henikoff, S. (2003) Spectrum of chemically induced mutations from a large-scale reverse-genetic screen in Arabidopsis. Genetics 164, 731-740. Abstract
Kulinski, J. Besack, D., Oleylowski, C.A., Godwin, A.K. and Yeung, A.T. (2000) CEL I enzymatic mutation detection assay. Biotechniques 29, 44-48.
** McCallum, C.M., Comai, L., Greene, E.A., and Henikoff, S. (2000) Targeted screening for induced mutations. Nat. Biotechnol. 18, 455–457. Abstract Library Entry
** McCallum CM, Comai L, Greene EA, Henikoff S (2000) Targeting induced local lesions in genomes (TILLING) for plant functional genomics. Plant Physiol. 123, 439–442 Abstract Library Entry
Oleykowski, C.A., Bronson Mullins, C.R., Godwin, A.K. and Yeung, A.T. (1998) Mutation detection using a novel plant endonuclease. Nucleic Acids Res. 26, 4597-4602. Abstract
Perry, J.A., Wang, T.L., Welham, T.J., Gardner, S., Pike, J.M., Yoshida, S. and Parniske, M. (2003) A TILLING reverse genetics tool and a Web-accessible collection of mutants of the legume Lotus japonicus. Plant Physiol. 131, 866-871. Abstract
Qiu, P. Shandilya, H., D’Alessio, J.M., O’Connor, K., Durocher, J. and Gerard, G.F. (2004) Mutation detection using Surveyor nuclease. Biotechniques 36, 702-707. Abstract
Qiu, P., Shandilya, H. and Gerard, G.F. (2005) A method for clone confirmation using a mismatch-specific DNA endonuclease. Mol. Biotechnol., 29, 11-18. Abstract
Scaffino, M.F., Pilotto, A., Papadimitriou, S., Sbalzarini, M., Ansaldi, S., Diegoli, M., Porcu, E., Grasso, M., Brega, A. and Arbustini, E. (2004) Heteroduplex detection with a plant DNA endonuclease for standard gel electrophoresis. Transgenics 4, 157-166.
Sokurenko, E.V. (2001) Discovering the sweeping power of point mutations using a GIRAFF. Trends Microbiol. 9, 522-525. Abstract
Sokurenko, E.V., Tchesnokova, V., Yeung, A.T., Oleykowski, C.A., Trintchina, E., Hughes, K.T., Rashid, R.A., Brint, J.M., Moseley, S.L. and Lory, S. (2001) Detection of simple mutations and polymorphisms in large genomic regions. Nucleic Acids Res. 29, e111. Abstract
Till, B.J., Reynolds, S.H., Greene, E.A., Codomo, C.A., Enns, L.C., Johnson, J.E., Burtner, C., Odden, A.R., Young, K., Taylor, N.E., Henikoff, J.G., Comai, L. and Henikoff, S. (2003) Large-scale discovery of induced point mutations with high-throughput TILLING. Genome Res. 13, 524-530. Abstract
Yang, B., Wen, X., Kodali, N.S., Oleykowski, C.A., Miller, C.G., Kulinski, J., Besack, D., Yeung, J.A., Kowalski, D. and Yeung, A.T. (2000) Purification, cloning, and characterization of CEL I nuclease. Biochemistry 39, 3533-3541. Abstract

View the online version of our Mutation Discovery and PCR Reference Handbook.