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TOROGreen® Blood Direct PCR Kit
    Publish time 2019-09-25 08:48    

TOROGreen® Blood Direct PCR Kit is is designed to amplify DNA directly from whole blood. No DNA purification is required prior to PCR. The kit is based on engineered hotstart KOD DNA Polymerase, a extremely robust PCR enzyme,which can tolerant of many PCR inhibitors present in blood and retains its activity at blood concentrations of up to 40% in the PCR reaction. TOROGreen® 6×DNA loading buffer contains the fluorescence dye, which is directly detected on 470nm LED transillum inator after the DNA electrophoresis. 

DESCRIPTION

TOROGreen® Blood Direct PCR Kit is is designed to amplify DNA directly from whole blood. No DNA purification is required prior to PCR. The kit is based on engineered hotstart KOD DNA Polymerase, a extremely robust PCR enzyme,which can tolerant of many PCR inhibitors present in blood and retains its activity at blood concentrations of up to 40% in the PCR reaction. TOROGreen® 6×DNA loading buffer contains the fluorescence dye, which is directly detected on 470nm LED transillum inator after the DNA electrophoresis. 

FEATURES

-Easy-to-usesample is added directly to PCR reaction, therefore there is no need for time-consuming and expensive DNA purification steps.
-Widely applicable—allows analysis of blood samples with different preservatives (e.g., heparin, EDTA, citrate), storage conditions and from various species..

-High performance: can amplify 40kb genomic DNA and 90% GC-rich target with 80-fold higher fidelity than Taq DNA polymerase. 

APPLICATION

- Transgene detection

- Genotyping
- Knockout analysis
- Sequencing

COMPONENTS

The kit includes the following reagents, which can be used for 100 reactions for a total 20ul reaction volume.

   Cat No. :DPK-400

   2× ASFastTM Direct PCR Mix                               1ml ×1tube

   TOROGreen®  6×DNA loading buffer                1ml ×1tube

PROTOCOL

1.Standard reaction setup.

1Prepare the mixture on ice as the following components.


  

 

Notes:

-Primers should be 22-35 bases with  Tm 65°C.

-Optimal primer concentration is 0.3uM. In the case of long targets (10 kb), reduced primers concentration (0.15uM) may give more effective amplification.

-When PCR yield is low, increased primers concentration (0.5uM) may give more effective amplification.

-With 2× ASFastTM Direct PCR Mix, it is possible to use a wide range of blood concentrations in the reaction (from 2.5% to 25%, or in some cases even up to 40%). The recommended starting point is 2.5%. In general, if higher blood percentage (>10%) is used, higher reaction volume (up to 50 μL) is recommended, and some optimization may be required (in particular the MgSO4 concentration may need to be increased up to 4.5 mM final concentration). For small amplicons, whole blood up to 40% can be used.

 

3.Set up cycling conditions

 

Notes:

-Poor amplification may be improved by changing the denaturation step to 95°C, 15sec.

4.Electrophoresis.

-Add 1 volume of TOROGreen® 6×DNA loading buffer to 5 volumes of PCR products.

-Mix well, spin down and load 5ul of the mixture and 2ul of the suitable TOROGreen® Loading Marker.

-Run on agarose electrophoresis to detect PCR products and marker .No additional dye is required for the PCR products and marker.

STORAGE
     This reagent  his reagent can be stored at 4°C for 1 months. For longer storage,  this reagent should be kept at -20°C


Ordering Information


Cat.No.


Product Name


Size


Store at


Price


Data Sheet


DPK-400


TOROGreen®Blood  Direct PCR Kit 


20μL×100 Reactions

 

-20°C


US$     100.00



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