Russian translation: раствор для переноса [продуктов ПЦР] на мембрану
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For the spotting and binding of DNA-probes onto QMT Epoxy Slides and QMT Aldehyde Slides we recommend the optimized QMT Spotting Solution I, II or III. For QMT Amino Slides Spotting Solutions I, IV and V are recommended. Make your own decision by selecting the best spotting solution appropriate and optimize your experiments and your spotting setup.
Why five different spotting solutions?
We recommend to compare the results of a complete immobilization and hybridization procedure with all 5 spotting solutions compared with e.g. 3x or 5x SSC. The results always depend on your equipment, your probes and your experimental conditions.
* Spotting Solution I is recommended to create relatively small spots with pin and pipetting spotting systems.
* Spotting Solution II shows good results using the Ring-And-Pin Technology.
* Spotting Solution III is recommended to create larger spots compared to solution I with pin and pipetting spotting systems.
* Spotting Solutions IV and V are designed to yield high intensity spots on Amino slides. They are not intended for usage on Epoxy- and Aldehyde slides.
The probes should be dissolved in water and diluted 1:1 with 2x QMT Spotting Solution to yield a 1x spotting solution.
Therefore, the DNA-molecules have to be adjusted to the double of the final spotting concentration according to the recommended values in the following table. For quantitative validations, e.g. gene expression profiles, the DNA-molecules to be immobilized must always be available in excess.
Low-density microarrays that utilize short oligos (<100 nt) for capture are highly attractive for use in diagnostic applications, yet these experiments require strict quality control and meticulous reproducibility. However, a survey of current literature indicates vast inconsistencies in the spotting and processing procedures. In this study, spotting and processing protocols were optimized for aldehyde-functionalized glass substrates. Figures of merit were developed for quantitative comparison of spot quality and reproducibility. Experimental variables examined included oligo concentration in the spotting buffer, composition of the spotting buffer, postspotting “curing” conditions, and postspotting wash conditions. Optimized conditions included the use of 3–4 μM oligo in a 3× standard saline citrate/0.05% sodium dodecyl sulfate/0.001% (3-[(3-cholamidopropyl) dimethylammonia]-1-propane sulfonate) spotting buffer, 24-h postspotting reaction at 100% relative humidity, and a four-step wash procedure. Evaluation of six types of aldehyde-functionalized glass substrates indicated that those manufactured by CEL Associates, Inc. yield the highest oligo coverage.
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раствор для переноса [продуктов ПЦР] на мембрану
раствор для переноса олигонуклеотидов на мембрану
Natalie Poland Local time: 20:24 Specializes in field Native speaker of: Russian PRO pts in category: 3905