点赞

收藏

分享

Mag-Bind Blood DNA Maximum Yield Protocol (2ml-6ml)

互联网2013-11-13

826

实验试剂

1. Absolute ethanol (96%-100%)

实验设备

1. Nuclease-free 50 ml centrifuge tube

2. Water bath, incubator or heating block preset at 65°C

3. Magnetic separation device for 50 ml tube

实验步骤

1. Place a empty, uncapped 50 ml conical tube into a 50ml tube holder.

2. Add 1-6 ml blood sample to 50ml tube.

3. Add indicated volume of PBS, MSL, Proteinase K and RnaseA showed in the table to the sample. Mix throughly by vortexing

4. Incubate sample at 65°C for 20 min. Briefly vortex the tube few times during incubation. Cool the sample to room temperature by incubating at room temperature for 5 minutes.

5. Add indicated volume of absolute ethanol showed in the table to the lysate. Mix throughly by inverting or voretxing.

6. Add 300ul Mag-Bind Particles Solution C to the sample. Mix gently by inverting, shaking or pipetting up and down 20 times. Incubate at room temperature for 10 minutes.

Note: Complete resuspension of cell pellet is vital for obtaining good DNA yields.

7. Place the tube on a magnetic separation device to magnetize the magnetic particles. Lysate will clear when the Mag-Bind particles have completely moved toward the ，magnet.

8. Completely remove and discard the cleared supernatant. Remove any droplets of liquid from the wall of the tube with the pipettor.

9. Remove the tube containing the Mag-Bind particles from the magneti c separation device. Add 5 ml Buffer MP/Ethanol Mixture to the sample.

Note: MP/Ethanol mix has to be prepared freshly.

10. Resuspend the Mag-Bind particles pellet by vortexing. Incubate 3 minutes at room temperature. During incubation, mix the sample several times by vortexing or pipetting up and down. Complete resuspension of the Mag-Bind particles pellet by pipetting up and down or vortexing is critical to obtain good results.

11. Transfer the sample to a new 50 ml tube and Incubate the sample at room temperature for 5 minutes.

12. Place the tune onto the magnetic separation device to magnetize the Mag-Bind particles.Completely remove and discard the cleared supernatant.

13. Remove the tube containing the Mag-Bind particles from the magnetic separation device. Add 10ml SPM Buffer to the sample.

14. Resuspend the Mag-Bind particles pellet by vortexing. Incubate 3 minutes at room temperature. Complete resuspension of the Mag- Bind particles pellet is critical to obtain good results.

15. Place the tube onto the magnetic separation device to magnetize the magnetic particles. Completely remove and discard the cleared supernatant.

16. Optional: Wash magnetic particles with SPM by repeating step 13-15.

17. Leave the tube to air dry on the magnetic separation device for 5 minutes. Remove any residue liquid with a pipettor. Do not over dry the pellet.

18. Remove the tube from the magnetic separation device. Add 2.5 ml Elution Buffer or water to elute DNA from the Mag-Bind particles. Mix throughly by vortexing for 30 seconds. Incubate 10 minutes at 65°C.

19. Place the tube onto a magnetic separation device to magnetize the Mag-Bind particles.

20. Transfer the cleared supernatant containing purified DNA to a new 1.5 ml tube.

相关产品推荐

FastStar DNA Polymerase，阿拉丁

FastStar DNA Polymerase，阿拉丁

￥747.90

ML604086，850330-18-6，阿拉丁

ML604086，850330-18-6，阿拉丁

￥761.90

Bind-Silane 亲和硅烷

Bind-Silane 亲和硅烷

￥188

Mag-Fluo-4 AM，1097709-31-3，阿拉丁

Mag-Fluo-4 AM，1097709-31-3，阿拉丁

￥3600.90

KTO: KIT A643 VALIDATION PROTOCOL (L)-Anatel A643a Accessories

KTO: KIT A643 VALIDATION PROTOCOL (L)-Anatel A643a Accessories

￥50094

相关问答

问

有能装下 500ml 样品的透析袋吗？

问

WB显出膜没有条带

问

质粒 DNA 比基因组 DNA 更大吗？

1 回答 2848 围观

相关方法

差异 DNA 的 PCR 扩增实验

2024-05-14

技术和方案3 酵母 DNA 分离

2024-05-14

利用 Polybrene 进行 DNA 转染实验

2024-05-14

推荐阅读

Mag-Bind Blood DNA Maximum Yield Protocol (7ml-10ml)

SQ Blood DNA Maxi Protocol for 4-10 ml whole blood

E.Z.N.A. Mag-Bind Blood DNA Protocol (1-100 ul Blood)

提问

扫一扫

丁香实验小程序二维码

实验小助手

丁香实验公众号二维码

扫码领资料

反馈

TOP

打开小程序