全血基因组DNA小量快速提取试剂盒-常用生化试剂-试剂-生物在线
全血基因组DNA小量快速提取试剂盒

全血基因组DNA小量快速提取试剂盒

商家询价

产品名称: 全血基因组DNA小量快速提取试剂盒

英文名称: Whole Blood DNA mini

产品编号: 116 505

产品价格: 0

产品产地: 进口

品牌商标: VWI

更新时间: null

使用范围: null

杭州莱枫生物科技有限公司
  • 联系人 :
  • 地址 : 杭州市江干区笕丁路168号
  • 邮编 :
  • 所在区域 : 浙江
  • 电话 : 159****9846
  • 传真 : 0571-56287646
  • 邮箱 : 185121481@qq.com

全血DNA小量快速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.快速、简单、高效,整个过程只需要10多分钟;

2.无需蛋白酶K裂解;

3.无需水浴加热;

4.无需分离白细胞;

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;

                                       Kit Contents

Whole blood DNA mini Kit

10

50

500

Cat No.

116 505-S

116 505

116 506

DNA binding columns

10

50

500

Colletion Tubes

10

   50

500

Buffer AC1

6ml

30ml

100ml×3

Buffer AC2

1.2ml

6ml

60ml

Buffer W1

4ml+2.5ml

19ml +12ml ethanol

80ml +50ml ethanol×2

Buffer W2

4ml+9ml

19ml +46ml ethanol

100ml +30ml ethanol×5

Buffer TE

1ml

10ml

50ml

 

4.2 Purification of Tatol DNA from Whole Blood

                                                          

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

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Centrifuge

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

--------------------------------------------------------------------------------------------------------------------------------------                                                                                            12,000×g 5 min             

●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


                                                                                    


After 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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●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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●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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After 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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To 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 70increases 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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The microcentrifuge tube now contains the eluted DNA.


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