whole blood DNA mini全血基因组DNA小量**提取试剂盒

全血DNA小量**提取试剂盒适合从小于250ul体积的全血DNA中提取多至12ug的基因组DNA。

1.**、简单、*，整个过程只需要10多分钟；

2.*蛋白酶K裂解；

3.*水浴加热；

4.*分离白细胞；

5.裂解液直接裂解细胞（白细胞），并能灭活血液当中的有害细菌和病毒；有效避免操作者被感染的可能；

4.2 Purification of Tatol DNA from Whole Blood

                                                          

①To a nuclease free 1.5 ml microcentrifuge tube

●Add 500 μL ofBuffer AC1.

 

▲     Before starting the purification reaction,                                                                                     500 μLBufferAC1        

warm up the Elution Buffer(TE or H2O) to 70°C.                                

 

 

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②Add 200-250 μl of anti-coagulated whole blood. Close the cap of the Microfuge

tube and mix by vortexing at top speed for 10-30 seconds.

 

●If your sample volume is less than 200 μL, the sample volume should be

expanded with PBS .

●If using avian blood or amphibian blood, combine <10 μl of blood with 200 μl of PBS.                                200 μL Blood                

 

●To extract genomic DNA from clotted or dried blood, place the sample in a

mortar (ambient temperature) and add 200 μl of 20 mM Tris, 10 mM EDTA, pH 8.5.

Grind rapidly for 30 seconds to disperse the sample.

Add 500 μl of Buffer AC1 pre-heated to 50°C and grind briefly or pipette to

dissolve the sample. Transfer the sample to a 1.5 ml Microfuge tube with a

transfer pipette or other device. Vortex for 10 seconds to further dissolve the

dried or clotted blood.

 




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③Add 100 μl of Buffer AC2 and mix byvortexingat top speed for 30 seconds.                                    100 μLBufferAC2

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④Centrifuge

12,000×g for 5-10 minutes at ambient temperature to pellet cellular debris.

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⑤●Insert one DNA binding column in one Collection Tube.                    

●Pipet the sample into the upper buffer reservoir of the DNA binding column.

●Insert the DNA binding column assembly into a standard tabletop centrifuge.

●Centrifuge 1 min at 12,000×g.

 




4.3 Protocol for Washing and Elution




                                                                                    




①After centrifugation:




●Remove the DNA binding column from the Collection Tube, discard the flowthrough liquid.                                              500ul W1




●Combine the DNA binding column with the Collection Tube.




●Add 500 μLBuffer W1. Removal Buffer to the upper reservoir of the Filter Tube.                                                                 12,000×g 1min




●Centrifuge 1 min at 12,000×g.




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②●Remove the DNA binding column from the Collection Tube, discard the flowthrough liquid.                                           500ul W2




●Combine the DNA binding column with the Collection Tube.




●Add 500μLBuffer W2to the upper reservoir of the DNA binding column.                                                                                  12,000×g 1min            




●Centrifuge 1 min at 12,000×g and discard the flowthrough.




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③●Remove the DNA binding column from the Collection Tube, discard the flowthrough liquid.                                         700ul W2        




●Combine the DNA binding column with the Collection Tube.




●Add 500 μLBuffer W2to the upper reservoir of the DNA binding column.                                                                               12,000×g 1min




●Centrifuge 1 min at 12,000×g and discard the flowthrough.




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④After discarding the flowthrough liquid:




●Centrifuge the DNA binding column for additional 1 min at full speed.                                                                                          12,000×g 1min




●Discard the Collection Tube.




▲The extra centrifugation time ensures removal of residual Wash Buffer.




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⑤To elute the DNA:




●Insert the DNA binding column into a clean, sterile 1.5 ml microcentrifuge tube.




●Add 200 μL prewarmedBuffer TEto the upper reservoir of the DNA binding column.




●Centrifuge the tube assembly for 1 min at 12,000×g 1min.




▲Elution with 50μL (instead of 200μL) increases the final DNA concentration in the eluate ,                                             12,000×g 1min




but also decreases the overall DNA yield.




▲Warm up theBuffer TEto 70℃increases the final DNA concentration in the eluate.




▲If you wish to remove RNA from the eluted DNA treat your sample as follows:




Add to the RNase A 2μL (10mg/ml) and incubate as appropriate(37℃,15min).




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⑥The microcentrifuge tube now contains the eluted DNA.




Either use the eluted DNA directly or store the eluted DNA at +2 to +8℃or15 to 25℃for later analysis.




                                                        

 

 

50次     260 元

500次   2400元