Differential Display DDRT-PCR Custom Service
Identification and Isolation of Modulated Genes



Differential Display is a powerful technique for identifying and isolating genes that display modulation under a given stimulus or environmental perturbation. The DDRT-PCR Kit from Display Systems Biotech provides a comprehensive protocol and complete set of reagents to perform this process in your own lab. Despite the widespread acceptance of the DDRT-PCR Kit as a gene discovery tool, the protocols remain labor-intensive and can take several weeks to obtain the desired results. Because Display Systems Biotech performs DDRT-PCR on a routine basis you can save time and avoid pitfalls inherent in the complex procedure (that often lead to a high level of false positives) by allowing us to perform the procedure for you; we guarantee results. You send us the RNA (or we'll isolate it for you), we reverse transcribe it and amplify the cDNA from each sample in duplicate. The amplified products are separated on a sequencing gel and candidate sequences are reamplified from gel slices, confirmed by SSCP and/or RPA, cloned, sequenced and analyzed. We can do some or all of the process as an interaction between customer and laboratory. You are in the driver's seat throughout the process

Flow of Events

  1. Cells or tissue to be compared are provided by customer;
  2. RNA is extracted from each sample and DNase-treated;
  3. A subpopulation of cDNA is synthesized using an anchored primer. A different subpopulation of cDNA is made with each anchored primer used.
  4. The cDNA subpopulation is amplified and the results are separated on a 48 lane sequencing gel (4 x 12 PCR reactions for one control and one test sample, in duplicate);
  5. Bands that identify up or down regulation are excised from the gel and the sequences are reamplified.
  6. Reverse Northern or SSCP analysis is performed to confirm modulation;
  7. Sequencing and analysis of reamplified sequences;

    8. Optional:

  8. Northern blot or ribonuclease protection analysis;
  9. Library screening for full-length clones;
  11. Sequencing of full-length clone;
  12. Computer analysis of sequence.







Two-Hybrid Characterization Screens

Differential Display Custom Service
Components
DSB800-101RNA Extraction (regular)$190.00
DSB800-102RNA Extraction (difficult)$430.00
DSB800-103DNase Treatment$45.00
DSB800-104cDNA synthesis:1 anchored primer; PCR of cDNA populations using upstream and anchored primer in duplicate$3000.00
DSB800-105-112Additional anchored primer 1-9$1600.00
DSB800-1141st added sample, DSB800-104$1125.00
DSB800-115ea. added sample, DSB800-104$750.00
DSB800-1161st added sample, DSB800-105:112$600.00
DSB800-117ea. added sample, DSB800-105:112$400.00
DSB800-118Identification of induced and reduced PCR bands and reamplification per sample$85.00
DSB800-119Northern/SSCP analysis, set-up fee$350.00
DSB800-120Northern/SSCP analysis/sample$280.00
DSB800-121Northern or RPA analysis/sample$800.00
DSB800-122Northern or RPA analysis/sample$150.00
DSB800-123Sequencing of reamplified sequence$1.50/bp
DSB800-124Library screening for full-length clonesinquire
DSB800-125Computer analysis of sequenceinquire
DSB810-100Point Mutation Analysisinquire
DSB820-100"Gene cloning, high level protein inquire expression and purification of the functional protein"inquire




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