For mRNA Differential Display with three one-base anchored oligo-dT primers and rationally designed arbitrary 13mers. These kits feature the latest generation of isotopic mRNA Differential Display technology (U.S. Patents 5,262,311; 5,599,672; 5,665,547; 5,965,409 and pending patents). Each kit contains all the reagents needed for differential display except RNA samples, Taq DNA polymerase, and -[33P] dATP. The MessageClean® is also highly recommended prior to differential display to remove trace amounts of DNA contamination from RNA samples. The major improvements over the previous RNAmap technology are: (1) Optimized primer length ensures excellent reproducibility and maximized coverage of expressed genes. (2) One-base anchored primers minimize the redundancy of the displayed cDNA and simplify the reverse transcription reactions. (3) Attached HindIII site to the primers facilitates the manipulation of the cDNA after cloning. See Technical Notes Section for further discussion on primer design, calculation of coverage, comparison of DD to other methodologies, and false positives. All kits are shipped on dry ice via overnight delivery. A detailed step-by step protocol is included. References: Liang, P. et al.: Differential display using one-base anchored oligo-dT primers. Nucleic Acids Res. 1994, 22:5763-5764. Ikeda, S. et al.: An aquaporin-like gene required for the Brassica self-incompatibility response. Science. 1997, 276:1564-1566. Taylor, G.A. et al: Identification of a Novel GTPase, the Inducibly Expressed GTPase, That Accumulates in Response to Interferon [gamma]. J. Biol. Chem. 1996, 271:20399-20405. More Publications by GenHunter customers Schematic representation of mRNA Differential Display using RNAimage® Kits. Three one-base anchored oligo-dT primers with 5' HindIII sites are used in combination with a series of arbitrary 13mers (also containing 5' HindIII sites) to reverse transcribe and amplify the mRNAs from a cell. Note the primer selectivity is provided by the 3' bases. See Technical Notes for more detailed information on the optimal primer designs for differential display. Differential Display using an RNAimage® Kit. Four RNA samples from rodent fibroblast (One normal, three tumorigenic) were compared by differential display using three different one-base anchored oligo-dT primers in combinations with three arbitrary 13mers. Note the excellent reproducibility and primer selectivity. The arrow indicates a candidate for tumor-specific gene expression.   Cat. No. RNAimage Kit 1 G501 RNAimage Kit 2 G502 RNAimage Kit 3 G503 RNAimage Kit 4 G504 RNAimage Kit 5 G505 RNAimage Kit 6 G506 RNAimage Kit 7 G507 RNAimage Kit 8 G508 RNAimage Kit 9 G509 RNAimage Kit 10 G510 RNAimage Kits 2-10 contain all the components of Kit 1, but each has 8 different arbitrary 13mers (H-AP9 to H-AP80). Choosing Kits: One could start with any RNAimage Kit since the sequences are random and designed to be maximally different from one another, but to ensure a maximum coverage, multiple kits may be needed (See Technical Notes). Individual components for the RNAimage Kits sold separately Description Cat. No. Volume H-T11G Anchor Primer (2µM) H-101-G 300 µL H-T11A Anchor Primer (2µM) H-101-A 300 µL H-T11C Anchor Primer (2µM) H-101-C 300 µL H-T11N Anchor Primer Set (2µM each) H-101-S 300 µL ea. H-AP Arbitrary Primers (2 µM each) H-AP1 to H-AP80 150 µL H-AP Primer Set 1 (H-AP1 to H-AP8) H-AP-A 150 µL H-AP Primer Set 2 (H-AP9 to H-AP16) H-AP-B 150 µL H-AP Primer Set 3 (H-AP17 to H-AP24) H-AP-C 150 µL H-AP Primer Set 4 (H-AP25 to H-AP32) H-AP-D 150 µL H-AP Primer Set 5 (H-AP33 to H-AP40) H-AP-E 150 µL H-AP Primer Set 6 (H-AP41 to H-AP48) H-AP-F 150 µL H-AP Primer Set 7 (H-AP49 to H-AP56) H-AP-G 150 µL H-AP Primer Set 8 (H-AP57 to H-AP64) H-AP-H 150 µL H-AP Primer Set 9 (H-AP65 to H-AP72) H-AP-I 150 µL H-AP Primer Set 10 (H-AP73 to H-AP80) H-AP-J 150 µL Glycogen (10 mg/mL) S301 500 µL 10X PCR Buffer S201 500 µL 5X RT Buffer S401 500 µL MMLV Reverse Transcriptase (100 unites/µL) S402 40 µL DNA Loading Dye S403 1 mL dNTP (250 µM) S501 200 µL dNTP (25 µM) S502 500 µL dH2O DH20 1.2 mL

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