2x Blood Direct PCR Master Mix
It contains Hotsar DNA Polymerase, dNTPs, Mg2+, Stabilizer, PCR enhancer and optimized buffer.
This direct PCR mix is easy and ready to use, can amplify blood DNA without any purification. Can amplify the High GC template DNA and oligos effectively.
This product has passed the following quality control assays: functional absence of double- and single-stranded endonuclease activity; >90% homogeneous by SDS gel electrophoresis.
Blood direct PCR
Store at -20°C
Reaction Mixture Set Up
|Component||50μl reaction||20μl reaction||Concentration|
|2×Blood Direct PCR Mix||25 μl||10 μl||as required|
|Forward Primer (10 μM)||1 μl||0.4 μl||0.2-0.4 μM each|
|Reverse Primer (10 μM)||1 μl||0.4 μl||0.5-0.4 μM each|
|ddH2O to final volume||50μl||20μl||Not applicable|
Note: We recommend take ≤10% whole blood as the template DNA. Gently mix well all the component with the vortex mixer after adding blood.
Primer concentration 0.1-1.0 μM As a reference to the setting range. When low Amplification efficiency can increase Primer concentration, If there is non-specific amplification can reduce the primer concentration, thus optimizing the reaction system.
Recommended thermal cycling conditions
94°C 2 min
94°C 30 sec
55-60°C 30 sec, 35-40 cycles
72°C 30-60 sec
1) In general, the annealing temperature is 5°C lower than primer TM. If can not get the ideal amplification efficiency, can low down annealing temperature; If there is the non-specific amplification can increase annealing temperature
2) The extension time should be set according to the size of the amplified fragment. The amplification efficiency of the hot-start DNA Polymerase in the mix is 1-2 kb / min.
3) The number of cycles can be set according to the downstream application of the amplification product. If the number of cycles less, the amount of amplification is not sufficient; if the number of cycles is too large mismatch probability will increase, non-specific background is serious. Therefore, the premise of ensuring the yield of products should minimize the number of cycles.
no repeated freezing and thawing and split charging for good amplification.