Telomeres and Telomerase: Methods and Protocols (Methods in by John A. Double, Michael J. Thompson PDF
By John A. Double, Michael J. Thompson
ISBN-10: 089603657X
ISBN-13: 9780896036574
John A. Double and Michael J. Thompson have accumulated a severely very important sequence of novel and crucial ideas for learning telomeres and telomerase. those easily reproducible equipment supply state of the art instruments to spot, degree, and research telomeres, to figure out telomerase expression on the RNA point, to figure out telomerase job, and to realize strength modifiers of this job. The thoughts for assaying telomerase task variety from ordinary radiological capture assays to nonradioactive equipment, from non PCR-based ways to options utilizing real-time PCR. Telomeres and Telomerase: tools and Protocols offers the center array of effective suggestions wanted this present day to boost telomerase inhibitors or diagnostic/prognostic telomerase markers.
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Extra info for Telomeres and Telomerase: Methods and Protocols (Methods in Molecular Biology Vol 191)
Sample text
Additional salt is not required for efficient precipitation. 20 Scherthan 7. Spin for 30 min at high speed. 8. Discard supernatant and wash pellet once with 70% EthOH. 9. Air-dry pellet by placing open tube for 5 –10 min in an incubator or drying block at 65°C. 10. Dissolve pellet in 16 µL TE buffer. 2. Dot Blot Test for Efficacy of Probe Labeling The dot blot test is applied to test the efficiency of the labeling reaction (see Note 6). 1. Spot 6 drops of 9 µL 6× SSC on a piece of parafilm. 2. ; Step 3) to the first drop, mix by repeated pipetting in and out of the drop.
5. Perform hybridization, washing, and detection steps of the WCP probe according to the manufacturer’s instructions. 4. 1. Image Acquisition Before image acquisition, check that the lamp of the microscope yields a field as homogeneous as possible. It is necessary to perform shading correction (5) with a image of flat-field to correct uneven illumination and a dark current image to detect the presence of hot pixels and noise attributable to CCD reading. 1. Perform the image acquisition with an anti-IR filter, which yields better contrasted images than without filter (see Note 7).
Dehydrate in ascending ethanols baths. 5. Perform hybridization, washing, and detection steps of the WCP probe according to the manufacturer’s instructions. 4. 1. Image Acquisition Before image acquisition, check that the lamp of the microscope yields a field as homogeneous as possible. It is necessary to perform shading correction (5) with a image of flat-field to correct uneven illumination and a dark current image to detect the presence of hot pixels and noise attributable to CCD reading. 1.
Telomeres and Telomerase: Methods and Protocols (Methods in Molecular Biology Vol 191) by John A. Double, Michael J. Thompson
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