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RNAimage Kits

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:
  1. Liang, P. et al.: Differential display using one-base anchored oligo-dT primers. Nucleic Acids Res. 1994, 22:5763-5764.

  2. Ikeda, S. et al.: An aquaporin-like gene required for the Brassica self-incompatibility response. Science. 1997, 276:1564-1566.

  3. 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
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.
Schematic
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.

Components
 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
DescriptionCat. No.Volume
H-T11G Anchor Primer (2ÁM)H-101-G300 ÁL
H-T11A Anchor Primer (2ÁM)H-101-A300 ÁL
H-T11C Anchor Primer (2ÁM)H-101-C300 ÁL
H-T11N Anchor Primer Set (2ÁM each)H-101-S300 ÁL ea.
H-AP Arbitrary Primers (2 ÁM each) H-AP1 to H-AP80150 ÁL
H-AP Primer Set 1 (H-AP1 to H-AP8)H-AP-A150 ÁL
H-AP Primer Set 2 (H-AP9 to H-AP16)H-AP-B150 ÁL
H-AP Primer Set 3 (H-AP17 to H-AP24)H-AP-C150 ÁL
H-AP Primer Set 4 (H-AP25 to H-AP32)H-AP-D150 ÁL
H-AP Primer Set 5 (H-AP33 to H-AP40)H-AP-E150 ÁL
H-AP Primer Set 6 (H-AP41 to H-AP48)H-AP-F150 ÁL
H-AP Primer Set 7 (H-AP49 to H-AP56)H-AP-G150 ÁL
H-AP Primer Set 8 (H-AP57 to H-AP64)H-AP-H150 ÁL
H-AP Primer Set 9 (H-AP65 to H-AP72)H-AP-I150 ÁL
H-AP Primer Set 10 (H-AP73 to H-AP80)H-AP-J150 ÁL
Glycogen (10 mg/mL)S301500 ÁL
10X PCR BufferS201500 ÁL
5X RT BufferS401500 ÁL
MMLV Reverse Transcriptase (100 unites/ÁL)S40240 ÁL
DNA Loading DyeS4031 mL
dNTP (250 ÁM)S501200 ÁL
dNTP (25 ÁM)S502500 ÁL
dH2ODH201.2 mL


 
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