ゴールドスタンダードの TaqMan MGB プローブケミストリを用いる Applied Biosystems TaqMan Copy Number Assays は、Applied Biosystems リアルタイム PCR 機器とソフトウェアを使用してゲノム DNA ターゲットのコピー数を測定します。TaqMan Copy Number Assays は、TaqMan Copy Number Reference Assay とともにデュプレックス qPCR に使用します。コピー数アッセイによりターゲット配列が検出され、リファレンスアッセイにより二倍体ゲノムにおいて 2 コピーで存在することが知られている配列が検出されます。その後、Applied Biosystems CopyCaller ソフトウェアを用いて、相対定量法により結果を解析します。
Custom Plus TaqMan Copy Number Assays は、ターゲットのヒト/マウスゲノム領域に適した設計済みアッセイがない場合の変異解析に最適なソリューションです。本アッセイは、無料の GeneAssist Copy Number Assay Tool を用いて、設計済みコレクションのアッセイの開発に使用されているのと同じバイオインフォマティクスパイプラインで設計されます。Custom Plus アッセイの設計プロセスには、事前のバイオインフォマティクス配列解析および in silico QC が含まれます。
1 tube containing a 20X (S and M sizes) or 60X (L size) mix of pre-formulated assay (1 probe and 2 primers).
Store at -15 to -25°C.
よくあるご質問(FAQ)
How can I reorder a Custom TaqMan Assay that I ordered previously?
To reorder the same Custom TaqMan Assay, you can follow these steps:
- Select an appropriate catalog number, considering the size of the assay that you want. You can find the different catalog numbers for Custom TaqMan Assays on their respective product pages on thermofisher.com.
- Once you have identified the necessary catalog number, go to www.thermofisher.com and click on "Quick Order", in the top right corner of the screen.
- In the following page, paste your selected catalog number in the "Catalog Number" field and paste the assay ID from your previous order in the "Assay ID" field.
- Click "Add to cart" and then in the following screen click "Proceed to check out".
Why do the values for the reference assay dye (VIC dye) vary for my TaqMan Copy Number Assay?
Please review the following possible causes and solutions:
-The reference sequence is not present or contains polymorphisms in the gDNA sample. Use an alternate copy number reference assay.
- There is variability in the amount of gDNA sample added to each reaction. Quantify the gDNA, then adjust the concentration of gDNA as needed.
Note: In general, the calculation of sample-level ΔCt accounts for variability in sample concentration.
- Pipetting was inaccurate. Check the pipette calibration of the pipettes, and pipette at least 5 µL of sample to prepare the reaction mix.
Why is the multicomponent signal for ROX dye not flat for my TaqMan Copy Number Assay?
Most likely incorrect dyes were selected for each target. Check the dyes selected for each target, then reanalyze the data.
Why was there a simultaneous increase in fluorescence from both the passive reference dye (ROX dye) and the reporter dyes for my TaqMan Copy Number Assay?
Most likely the sample evaporated. We recommend that you check the seal of the adhesive film for leaks before running the plate on the real-time PCR instrument.
Why was there a decrease in ROX dye fluorescence (passive reference dye) in my TaqMan Copy Number Assay?
Please review the following possible causes and solutions:
- There was insufficient Master Mix added to the reaction. Please follow accurate pipetting practices.
- Reagents are degraded. Ensure that the kits and reagents have been stored according to the instructions on the packaging and that they have not expired.