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ReadyMade™ Random Hexamers

Inventoried oligos comprising all possible sequences for a given length.

ReadyMade Random Hexamers can be used for various applications, including cDNA synthesis and identification of single nucleotide polymorphisms. The actual base composition confirmed by enzymatic degradation and HPLC. The synthesis method helps to ensure equal base representation.

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ReadyMade Random Hexamers (randomers) are oligonucleotides composed of all possible sequences for a given length. As the coupling efficiency for each individual phosphoramidite is slightly different, a special blend of all 4 bases at different molar ratios is used to achieve an even nucleotide representation. The batch is then tested by synthesizing test randomer oligos and confirming base composition by enzymatic degradation and HPLC. The use of randomers began in the early 1980s when random hexamers were employed in radiolabeling DNA probes [1,2].

A later application was the use of random sequence primers to detect random amplified polymorphisms (RAPDs) [3,4]. You can use random sequence oligonucleotides to identify single nucleotide polymorphisms (SNPs) as well as small scale chromosome events, primarily insertions or deletions [5,6].

Comparative genomic hybridization (CGH) has been developed to elucidate genome-wide sequence copy number variation (CNV) between different genomes, such as the differential amplification or deletion of genetic regions between tumor DNA and normal DNA from neighboring unaffected tissue [7-10]. These applications demand that the oligonucleotides have consistent base composition and minimal lot-to-lot variation.

 

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References

1. Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983;132(1):6-13. doi:10.1016/0003-2697(83)90418-9

2. Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Addendum. Anal Biochem. 1984;137(1):266-267. doi:10.1016/0003-2697(84)90381-6

3.Williams JG, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 1990;18(22):6531-6535. doi:10.1093/nar/18.22.6531

4. Forrest AK, Jarvest RL, Mensah LM, O'Hanlon PJ, Pope AJ, Sheppard RJ. Aminoalkyl adenylate and aminoacyl sulfamate intermediate analogues differing greatly in affinity for their cognate Staphylococcus aureus aminoacyl tRNA synthetases. Bioorg Med Chem Lett. 2000;10(16):1871-1874. doi:10.1016/s0960-894x(00)00360-7

5. Houldsworth J, Chaganti RS. Comparative genomic hybridization: an overview. Am J Pathol. 1994;145(6):1253-1260.

6. Lichter P, Bentz M, Joos S. Detection of chromosomal aberrations by means of molecular cytogenetics: painting of chromosomes and chromosomal subregions and comparative genomic hybridization. Methods Enzymol. 1995;254:334-359. doi:10.1016/0076-6879(95)54024-5

7. Kallioniemi A, Kallioniemi OP, Sudar D, et al. Comparative genomic hybridization for molecular cytogenetic analysis of solid tumors. Science. 1992;258(5083):818-821. doi:10.1126/science.1359641

8. Pinkel D, Segraves R, Sudar D, et al. High resolution analysis of DNA copy number variation using comparative genomic hybridization to microarrays. Nat Genet. 1998;20(2):207-211. doi:10.1038/2524

9. Emanuel BS, Saitta SC. From microscopes to microarrays: dissecting recurrent chromosomal rearrangements. Nat Rev Genet. 2007;8(11):869-883. doi:10.1038/nrg2136

10. Gresham D, Dunham MJ, Botstein D. Comparing whole genomes using DNA microarrays. Nat Rev Genet. 2008;9(4):291-302. doi:10.1038/nrg2335

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