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Technical Data Sheets

Gene Cone Chambers

EMS Catalog # 72390-96 & 72390-24

A Gene Cone Chamber encloses a 15mm diameter specimen area on a microscope slide to control evaporative loss of reagents during thermocycling. Reagents are pipetted into the top opening and top opening is sealed with an exclusive adhesive film that is impermeable to water vapor and accommodates pressure changes in the chamber.

1. Peel and place a self-adhesive Gene Cone Chamber over dry, or semi-dry specimen. Heat slide for 15 seconds at 95°C to set the adhesive to the slide. (Pressing with an inverted plastic 15ml culture tube or similar cylinder after the slide is heated, improves the adhesive seal around the perimeter of the specimen. The adhesive rim may over-lay portions of the tissue sections.)
2. Peel and place self-adhesive top seal onto the individual Gene Cone. The top seal may be applied without touching the adhesive by peeling and sticking one part of the seal at a time. (Note: The top seal is not designed to be removed. If you do peel the top seal off the chamber opening, press firmly on the seal with the slide.)
3. Remove the chamber after thermocycling by lifting the extended tab and peeling the chamber and adhesive off the slide. Proceed with detection of label and cellular analysis.

Using Gene Cone Chambers For In Situ DNA Amplification:

1. To denature DNA in the cells, pipette 60 - 100 ul distilled water into the chamber and heat at 95°C for 3-5 minutes without covering or until water has evaporated and before the cells dry out. After the initial denaturation of double-stranded template, 90 degree C denaturations are sufficient. If the chamber leaks, it is likely that the cycling denaturation temperatures are higher than needed.
2. To amplify DNA , bring slides to 60°C or the optimal annealing temperature. Add 50-100 ul reaction mix to chamber and close with self adhesive top seal.
   Note: Warm slides to 55-70°C before sealing top. Cycle.
3. To detect product directly, substitute one-tenth of the dTTP with no-isotopically labeled dUTP or one-tenth of the oligonucleotide primer with end-labeled primer. Less than 10 cycles are necessary when efficient amplification conditions are determined, even for single copy targets. Caution: Extended rounds of amplification may incorporate label indiscriminately. Overly cross-linked or overly protease-treated cells and tissue are less likely to amplify efficiently or retain morphology. Note: When removing chamber, adhesive remaining on treated slides (i.e., charged silanated) can be peeled off with forceps.
4. Temperature control. Either use a heating platen or oven calibrated for slides or cover a heating platen for tubes with a sturdy heat conducting material. Recommended: Set up in situ amplification cycling parameters for your thermocycler without using cells or tissue in the Gene Cone chambers. Use extracted DNA and reagents in the chambers which have previously given positive amplification results in tube reactions. Also when using a tube cycler with an internal temperature probe, program higher temperature setpoints in order to offset temperature of the slides on top of the heating platen. Program time periods comparable to what is used for solution reactions. Example:

Online Ordering

Gene Cone Chambers for in-situ DNA amplification and hybridization are available online from the EMS Catalog. For ordering or product information, click here.