![]() (Refer to the QuantStudio™ 12K Flex Real-Time PCR System Maintenance and Administration Manual, Appendix C, “Creating a custom dye plate”, for full details. Create a full plate of the dye using the selected concentration and run the custom dye calibration as normal, using 20 µL.Select the concentration to use by finding the dilution that will give you an acceptable signal in the following ranges:įor a 384-well plate: between 400,000 and 1,200,000įor a 96-well plate: between 1,400,000 and 4,300,000 When the run is complete, export and examine the raw data.Ensure that the filters of interest are selected. Alter the thermal profile so that it simply ramps to 60☌ with a 2 min hold. Set up a dummy run using the ‘Standard Curve’ option.Note: The volume would be the same for a 96-well or 384-well plate. In this example, sample concentrations would be 100, 200, 400, 800, and 1,600 (20 µL/well). Their ultrathin wall design will ensure rapid thermal transfer and a significant reduction in cycle and. A full plate is not needed see the example below. Primer design optimize your primer design using our custom oligo design tool. Get the basics on how to select the right PCR enzyme for your application. Use either the background buffer provided in the calibration kit or your own buffer to dilute the oligo. The most important considerations for any PCR reaction are: PCR enzymes enzyme selection can affect the yield and specificity of PCR products. For each custom dye, prepare samples in a concentration range of 100–2,000 nM, choosing a 2–3-fold difference in dilution points. followed by 14 cycles of touchdown PCR consisting of 15 s at 95C. Esco represents innovation and forward-thinking designs, which are all coupled with the highest standard quality since 1978. Amplifications using an Applied Biosystems thermal cycler (GeneAmp 2720) were. You will need an oligo sequence with the custom dye but without the quencher molecule. WELCOME TO ESCO Esco’s vision is to provide enabling technologies for scientific discoveries to make human lives healthier and safer. PC-controlled or run directly from touch screen The most important considerations for any PCR reaction are: PCR enzymes enzyme selection can affect the yield and specificity of PCR products. In a thermal cycler (The Applied Biosystems 2720 Thermal Cycler). Gene expression, genotyping, copy number variation, HRM, protein thermal shift, protein detection, mutation detection, miRNA, presence/absence, digital PCR 10 The genomic DNA was amplified using the Touchdown (TD) PCR assay11 and. 96-well (standard), 96-well (Fast), 384-well, TaqMan® Array Card, OpenArray® plate (4 plates)įAM™, SYBR®, VIC®, ROX™, NED™, TAMRA™ dyesġ0–100 µL (96-well standard) 15–30 µL (96-well Fast) 5–20 µL (384-well) ~1 µL (TaqMan® Array Card) 33 nL (OpenArray®Plate)Ħ excitation filters (450–670 nm), OptiFlex™ system (enhanced fluorescence detection), 6 emission filters (500–720 nm)įor Research Use Only. ![]()
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