Q：檢測準嗎？巢式 pcr 什么意思？為什么要做兩輪 pcr？

A：巢式 PCR 通過兩輪擴增的方式，增加了產(chǎn)品的特異性和靈敏度，可以對 10 拷貝的支原體進行檢測。

Q：GMyc-PCR 支原體檢測試劑盒，能測細胞嗎？細胞支原體測定方法是什么樣的？

A：可以的，如果使用細胞懸液做樣品，則需要提取DNA 后再進行 PCR。加入量一般為反應(yīng)體系 1/10 的量或更少。

Q：對于同一個樣品，為什么一步法恒溫試劑盒檢測結(jié)果為陽性，而 PCR 檢測為陰性呢？

A：一步法恒溫檢測試劑盒檢測的支原體的種類是 27 種，而 PCR 檢測方法檢測的是常見的 13 種支原體。樣品中污染的支原體可能不包括在 PCR 法檢測的支原體種類內(nèi)。

Q：巢式 PCR 法和一步法恒溫支原體檢測試劑盒他們的優(yōu)勢在哪些方面，怎么選擇？

A：PCR 法的靈敏度高于一步法檢測，而一步法的靈敏度是 10^3 個拷貝才能檢測出來。一步法檢測的細胞支原體種類達 20 多種多于 PCR 法。

Q：GMyc-PCR 支原體檢測結(jié)果如何判斷？

A：GMyc-PCR 支原體檢測方法通過巢式 PCR 的方式獲得檢測結(jié)果。整個實驗由 2 輪PCR 組成，檢測結(jié)果以第二輪為準。

Q：說明書中寫的是不加loading buffer，直接點樣就可以，我點樣之后所喲有樣品均沒有條帶（包含陽性對照），陽性對照加了loading buffeer才有條帶。

A： 經(jīng)確定客戶配膠的時候未加入EB染料，他們的染料是加在loading buffer中的，所以不加入自己的loading buffer跑不出條帶。

EB加入方式：

      1.如果配瓊脂糖凝膠的時候加入EB染料，跑膠的的樣品中不需要加入了

      2.如果配瓊脂糖凝膠的時候未加入EB染料，跑膠的的樣品中需要加入EB染料

Q：陽性對照是哪種支原體呢？

A：豬鼻子支原體

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