Test Label: Biotin labeled primer / FAM probeTest Format: AmplifyRP® XRT+
Xylella fastidiosa (Xf) is the causal agent of disease in multiple crops including Pierce"s disease in grape, citrus variegated chlorosis (CVC), almond leaf scorch disease and olive quick decline syndrome.Severe Xf infections can decimate entire crops and cause enormous financial losses to growers.
AmplifyRP XRT+ for Xf is an isothermal DNA amplification and detection system that offers unrivaled detection capabilities in an easy-to-use testing format.It is highly specific and offers equivalent, or better, sensitivity as published PCR methods while eliminating laborious and costly nucleic acid extractions.Prior molecular diagnostic experience is not required toperform AmplifyRP XRT+ tests.
XRT+ is our most flexible AmplifyRP platform and offers the option of either real-time detection (Image 2) or end-point detection (Image 3).Real-time detection is completed using the field deployable (battery operated) AmpliFire fluorometer.Assay parameters are loaded via barcode and results are automatically displayed as ( + ) or ( - ).
End-point detection can alternatively be completed by performing the amplification step in a small 42°C heat block followed by amplicon detection using an Amplicon detection chamber (sold separately).
Limit of Detection (LOD): 22 copies of Xf genomic DNA per reaction.
Specificity: No known cross-reactions to other pathogens
Total Assay Time: Less than 30 minutes when used with the AmpliFire as a real-time assay.Less than 45 minutes when used with the Amplicon Detection Chamber as an end-point assay.
Includes:
- XRT+ reaction pellets for Xf (48)
- PD1 Pellet diluent (55 mL)
- Amp1 extraction buffer (3 x 55 mL)
- Micro centrifuge tubes (50)
- Mesh extraction bags (50)
- User Guide
If you plan to use this assay as a real-time assay you will need to purchase the following items separately:
- AmpliFire Isothermal Fluorometer
If you plan to use this assay as a end-point detection assay you will need to purchase the following items separately:
- AmplifyRP Acceler8 starter pack
- Amplicon detection chambers
- Disposable transfer pipettes (optional if you don"t have laboratory pipettes)
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利用NheI和HindIII双酶切插入目的片段，为什么双酶切有目的片段和切开的质粒，测序却没有？
并且测序后在NheI位点后的序列变成了-GGCTAGGTAC-Kpn位点（CGTTTAAACTTAAGCTTG消失。之前和之后的序列都相符），之间的怎么都没了？
请教高人，这样的问题怎么解决？
多谢
你所说的“测序酶”在一般情况下就是“聚合酶”。
因为目前的的测序方法主要就是借助聚合反应。
当然，有些测序方法（比如曾经的SOLiD系统）是利用连接反应，那么它用的“测序酶”肯定是连接酶。
估计，也就是这个原因有人把它。
两个基因大小分别230kb和450kb，酶切后，450kb的有目的条带，但很弱，230kb的质粒条带亮，其下方有一很微弱条带，用的酶分别是：Ncol和Spel体系是（Takara，两个酶切体系不同，用的官网推荐体系）：
NcoI1μl
Spel1μl
10×KBuffer2μl
0.1%BSA2μl
DNA3ul
灭菌水upto20μl37℃4h电泳1h
期待着高手们的指点，谢谢。
【之前构建的4个重组质粒里也有两个出现这样的问题（aagtcc变成agtcc），但是酶切能切出插入的片段】
目前问测序公司得到的答复是样品可能不是单克隆，测序结果是不准确的，建议重新挑取单克隆测序。
不知道还有没有别的方法？
●应用酶水解多肽不会破坏氨基酸，也不会发生消旋化。水解的产物为较小的肽段。向左转|向右转
