• 1 x Oncomine Focus 检测、DNA Chef-Ready 检测板 (2X) • 1 x Oncomine Focus 检测、RNA Chef-Ready 检测板 (2X) • 2 x Ion AmpliSeq 试剂盒(用于 Chef DL8 试剂)
注:不包括用于 cDNA 合成的 SuperScript 转录酶。必须单独销售。
常见问题解答 (FAQ)
With the Oncomine Focus Assay, Chef-Ready Library kit, where can I find the combined barcoded AmpliSeq libraries when the run is complete?
After the completion of a run, the Ion Chef Instrument holds the barcoded libraries in the tube loaded in Position D of the Reagents cartridge. To avoid fluid loss due to evaporation, remove and cap the tube of combined barcoded libraries as soon as possible after run completion. Do not leave the tube in the instrument longer than 24 hours after the start of the run. After 24 hours from the start of the run, the instrument chiller will stop actively cooling, and the sample will be held at 27 degrees C.
The tube in Position D of the Ion AmpliSeq Chef Reagents DL8 Cartridge will contain 700 µL of combined barcoded libraries at completion of the run. The libraries are at >>100 pM (total combined library concentration) and are ready to use in template preparation. One can store unused portions of combined libraries at 4 to 8 degrees C for up to 1 month. For longer-term storage, store at -30 to -10 degrees C.
With the Oncomine Focus Assay, Chef-Ready Library kit, can I process fewer than 8 samples per Ion AmpliSeq run on the Ion Chef System?
Yes, fewer than 8 samples may be processed per run but keep in mind that a run consumes kit reagents for 8 samples regardless of the sample number.
If I process less than 8 samples using the Oncomine Focus Assay, Chef-Ready Library kit, do I need to quantitate the final combined output library?
If you are processing ≤5 samples, we recommend that you quantify your output combined library by qPCR to ensure that an optimal concentration is used in templating reactions.
If you are processing fewer than 8 samples, it is preferable to add replicates or positive control samples to the run. Otherwise, pipet 15 µL of nuclease-free water as non-template control into column 1 wells that do not contain a DNA sample.
Where should I add the AmpliSeq primer pools on the Ion Chef Instrument deck?
The primer pools should be added to the Reagent Cartridge on the Ion Chef Instrument in positions A and B. Tubes in Positions C and D can be left empty. Uncap all 4 tubes in positions A, B, C, and D in the Ion AmpliSeq Chef Reagents DL8 cartridge.
What is the RNA input for the Oncomine Focus Assay, Chef-Ready Library kit?
Each reverse transcription reaction requires 10 ng of DNase-treated total RNA (≥1.43 ng/µL).