1.Weigh 500 mg of glass beads in a 15 ml centrifuge tube, add 0.2-0.5 g soil sample. Add 1 ml Buffer SLX Mlus. Vortex at maximum speed for 3-5 minute to lyse sample. For the best result, A Mixer Mill,such as Fastprep-24,Mixer Mill 300, should be used.
称取500毫克的玻璃珠在15毫升离心管,添加0.2 -0.5 g土壤样品。加入1毫升SLX Mlus缓冲液。最大速度下搅拌3 - 5分钟直到样品溶解。为达到最佳效果,需要用到搅拌磨,如Fastprep-24、Mixer Mill 300。
2.Add 100 ul Buffer DS and vortex to mix.
加入100ulDS缓冲液，搅拌混匀。
3.Incubate at 70 ℃ for 10 minute. Briefly vortex the tube once during the incubation. For some difficult lysis bacterial. Increase the temperature to 90 ℃.
在70 ℃ 培养10分钟。在培养的过程中稍微地搅拌离心管一次，对一些比较难溶解的细菌，将温度调至90℃。
4.Centrifuge at 3000 rpm for 3 min at room temperature. Transfer 800 ul the supernatant into a new 2 ml tube and add 270 ul Buffer SP2.
Mix the sample throughly by vortexing.室温下3000rpm离心3分钟。转移800ul上清液到一个新的2 ml 管，然后加入270 ul SP2 缓冲液。搅拌，彻底地混匀样品。
5.Incubate in ice for 5 min. Centrifuge at full speed (13,000 ×g) in a microcentrifuge for 5 min at 4 ℃.
冰上孵育5分钟。在微离心机下，4℃全速（13,000 g）离心5分钟。
6.Carefully transfer supernatant to a new 2 ml tube and add 0.7 volume of isopropanol. Mix throughly by inverting tube for 20 -30 times. If the soil contains very low DNA, incubate the sample at -20℃ for 1 hour.
小心地转移上清液到一个新的 2 ml 管，然后加0.7体积的异丙醇。通过反向管彻底地混合20-30次。假如土壤中含少量的DNA，需要在-20℃ 孵育1h。
7.Precipitate DNA by centrifuge at full speed (13,000 ×g) for 10 minute at 4 ℃ .
4 ℃下全速(13,000 ×g)离心10分钟，沉淀DNA。
8.Carefully discard the supernatant and make sure not dislodge the DNA pellet. Invert the tube on a absorbent paper for 1 minute to drain the liquid. It is not necessary to dry the DNA pellet.小心地弃掉上清液，确保不搅动DNA沉淀。倒置离心管在吸水纸上1 分钟，使液体流尽。不需要干燥DNA沉淀。
9.Add 200 ul of Elution Buffer to the tube and vortex for 10 seconds.Incubate at 65℃ for 10- 20 minutes to dissolve the DNA pellet.加 200 ul 洗脱缓冲液，然后搅拌10秒钟。在 65℃ 孵育10-20 分钟，溶解DNA 沉淀。
10.Add 50 -100 ul of HTR Reagent. Mix throughly by vortexing for 10 minute. Note :completely resuspend HRT Reagent by shaking the bottle before use.
加 50- 100ul HTR 试剂，搅拌10分钟，彻底混匀。注意：用前摇动瓶子，充分悬浮HTR 试剂。
11.Incubate at room temperature for 2 minutes. Centrifuge at full speed (13,000 ×g) for 2 minutes.室温下孵育2分钟。全速 (13,000 ×g)离心2分钟。
12.Transfer cleared supernatant to a new2 ml tube.Note :if supernatant still have dark color from soil ,perform the HTR treatment again by repeat step 10-12.
转移清澈的上清液到一个新的2ml管。注意：假如上清液呈黑色，重复10-12步。
13. Add equal volume of XP2 Buffer ,mix by vortexing. For examper: if the sample from step 12 is 250 ul, add 250 ul Buffer XP2.
加入等量的XP2 缓冲液，搅动混匀。如：12步后的量是250ul，就加入250ul的XP2缓冲液。
14.Apply the sample from step 13 to a HiBind DNA Column assembled in a 2 ml collection tube(supplied).centrifuge at 10,000×g for 1 minute at room temperature. Discard flow-through liquid and re-use collection tube.
将13步的样加到组装于2ml收集管（已提供）的HiBind DNA Column。室温下10,000×g 离心1分钟，倒掉直流液体，再次使用收集管。
15.Place the column into the same 2 ml collection tube from previous step and add 300 ul XP2 Buffer. Centrifuge at 10,000×g for 1 minute. Discard the flow-through and collection tube.将柱子放到相同的先前的2ml收集管，然后加入3000ulXP2缓冲液。10,000×g离心1分钟。弃掉液体和收集管。
16.Place column into a new 2 ml collection tube(supplied) and wash by adding 700 ul SPW Wash Buffer diluted with absolute ethanol. Centrifuge at 10,000×g for 1 minute.Discard flow-through liquid and re-use collection tube in next step.Note:SPW Wash Buffer is provided as a concentrate and must bediluted with absolute ethanol as indicated on the bottle and page 4.
将柱子置于一个新的2ml收集管（已提供），加入700ul 用无水乙醇稀释的SPW洗脱液洗脱。10,000×g离心1分钟。弃掉直流液，收集管下一步再次使用。注意：SPW洗脱液必须浓缩，必须用无水乙醇再次稀释，此项注明在瓶子上和第4页。
17.Repeat step 16 with a second 700 ul SPW Wash Buffer.
重复16步。
18.Discard liquid and re-insert the column to the empty collection tube,centrifuge the column at full speed(13,000 ×g) for 2 minutes at room temperature. This step is critical in removing traces of ethanol that will interfere with downstream applications.
弃掉液体，重新插入柱子到空的收集管，室温下全速离心2分钟。这步对清除乙醇痕迹很重要。因为乙醇会干扰下面步骤的应用。
19.Place column into a clean 1.5 ml microcentrifuge tube (not supplied). Add 30- 100 ul Elution Buffer directly onto the centre of HiBind matrix. Incubate at 65 ℃ for 10-15 minutes.
将柱子置于一个干净的1.5ml的微离心机管（没有提供）。直接加入30-100ul洗脱液在HiBind matrix的中心。在65 ℃下孵育10-15分钟。
20.Centrifuge at full speed (13,000 ×g) for 1 min to Elute DNA.
全速离心1分钟，洗提DNA。
21.Repeat elution step 19-20 with a second 30-100 ul Elution Buffer.
重复洗脱步骤19-20。
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