基本描述

英文名称

Blood Genomic DNA Mini Kit (0.1-1 mL)

储存温度

2-8°C储存,室温

运输条件

冰袋运输

产品介绍

B665530	Component	50 T	200 T	Storage
B665530A	Buffer RCL	125 mL	2×260 mL	2-8℃
B665530B	Buffer GR	15 mL	50 mL	RT
B665530C	Buffer GL	15 mL	50 mL	RT
B665530D	Buffer GW1 (concentrate)	13 mL	52 mL	RT
B665530E	Buffer GW2 (concentrate)	15 mL	50 mL	RT
B665530F	Buffer GE	15 mL	60 mL	RT
B665530G	Proteinase K	1.25 mL	4×1.25 mL	RT
B665530H	Spin Columns DM with Collection Tubes	50 sets	200 sets	RT

产品简介

本试剂盒适用于从新鲜或冷冻的全血（用柠檬酸盐、EDTA或肝素等抗凝剂处理过的血液样品）、血浆、血清、血沉棕黄层、淋巴细胞、无细胞体液等样本中提取总 DNA，包括基因组DNA，线粒体DNA及病毒DNA。本品可以处理0.1-1 mL的全血，最高得率可达30 μg，可纯化获得大小为100 bp到50 kb的DNA，纯化的DNA产量高、质量好，最大限度去除蛋白、色素、脂类及其他抑制性杂质污染，可直接用于PCR、荧光定量PCR、酶切和Southern Blot等实验。

自备试剂：无水乙醇。

实验前准备及重要注意事项：

1．样品应避免反复冻融，否则会导致提取的DNA片段较小且提取量也下降。

2．本试剂盒最多可以提取0.1-1 mL全血样品或1×107个白细胞。

3．第一次使用前应按照试剂瓶标签的说明先在Buffer GW1和Buffer GW2中加入无水乙醇。

4．使用前请检查Buffer GL是否出现结晶或者沉淀，如有结晶或者沉淀，请将Buffer GL 于56℃水浴孵育重新溶解。

5．试剂盒中的Buffer RCL浑浊后不能继续使用。

操作步骤：

1．样品处理： 1a. 提取200 uL血液样品时，向离心管（自备）中加入样本后，可直接进行下一步实验。 1b. 当血液样本量小于200 μL时，加入Buffer GR补足至200 μL，再进行下一步实验。 1c. 当血液样本量超过200 μL时，加入1~2.5倍体积的Buffer RCL，轻轻涡旋或颠倒混匀， 12,000 rpm（~13,400×g）离心1分钟，小心吸弃上清液，如果沉淀中还有红色，可以重复以上步骤一次。然后向沉淀中加入200 μL Buffer GR，震荡至彻底混匀,再进行下一步实验。 1d. 如果处理血液样本为禽类、鸟类、两栖类或更低级生物的抗凝血，其红细胞为有核细胞，血液样本量为5-20 μL，可加入Buffer GR，补足至200 μl后进行后续实验。注意：如果下游试验对RNA敏感，可加入4 μL RNase A（100mg/mL）溶液，震荡15秒，室温放置5 分钟。RNase A本试剂盒并未提供，如需要可单独向本公司订购，货号：CW0601S。

2．向以上溶液中加入20 μL Proteinase K，混匀。

3．加入200 μL Buffer GL，震荡至彻底混匀。注意：不要将Proteinase K和Buffer GL进行预混。

4．56℃孵育10分钟，其间颠倒混匀数次。注意：孵育10分钟DNA的产量已经达到最大，继续延长孵育时间对DNA产量和纯度没有影响。

5．加入200μL无水乙醇，颠倒混匀数次。短暂离心，使管壁和壁盖上的液体集中到管底。

6．将步骤5所得溶液全部加入到已装入收集管的吸附柱(Spin Columns DM）中，若一次不能加完溶液，可分多次转入。12,000 rpm离心1分钟，倒掉收集管中的废液，将吸附柱重新放回收集管中。

7．向吸附柱中加入500 μL Buffer GW1（使用前检查是否加入无水乙醇）, 12,000 rpm 离心1分钟，倒掉收集管中的废液，将吸附柱重新放回收集管中。注意：如果提取样品是小鼠或猴子等血红素难以除去的种属的血液基因组，建议重复步骤7。

8．向吸附柱中加入500 μL Buffer GW2（使用前检查是否加入无水乙醇），12,000 rpm 离心1分钟，倒掉收集管中的废液，将吸附柱重新放回收集管中。注意：如需进一步提高DNA纯度，可重复步骤8。

9．12,000 rpm离心2分钟，倒掉收集管中的废液。将吸附柱置于室温数分钟，以彻底晾干。注意：这一步的目的是将吸附柱中残余的乙醇去除，乙醇的残留会影响后续的酶促反应（酶切、PCR 等）

10．将吸附柱置于一个新的离心管（自备）中，向吸附柱的中间部位悬空加入50-200 μL Buffer GE或灭菌水，室温放置2-5分钟，12,000 rpm离心1分钟，收集DNA溶液，-20℃保存DNA。注意：1）如果下游实验对pH值或EDTA敏感，可以用灭菌水洗脱。洗脱液的pH值对洗脱效率有很大影响，若用水做洗脱液应保证其pH值在7.0-8.5（可以用NaOH将水的pH值调到此范围），pH值低于7.0时洗脱效率不高。 2）如果要提高DNA的终浓度，可以将所得的DNA洗脱液重新加至吸附膜上，室温放置2-5分钟， 12,000 rpm离心1 分钟。 3）因为保存在水中的DNA会受到酸性水解作用的影响，如需长期保存，推荐用Buffer GE洗脱并于-20℃保存。

B665530	Component	50 T	200 T	Storage
B665530A	Buffer RCL	125 mL	2×260 mL	2-8℃
B665530B	Buffer GR	15 mL	50 mL	RT
B665530C	Buffer GL	15 mL	50 mL	RT
B665530D	Buffer GW1 (concentrate)	13 mL	52 mL	RT
B665530E	Buffer GW2 (concentrate)	15 mL	50 mL	RT
B665530F	Buffer GE	15 mL	60 mL	RT
B665530G	Proteinase K	1.25 mL	4×1.25 mL	RT
B665530H	Spin Columns DM with Collection Tubes	50 sets	200 sets	RT

Product Introduction
This reagent kit is suitable for extracting total DNA, including genomic DNA, mitochondrial DNA, and viral DNA, from fresh or frozen whole blood (blood samples treated with anticoagulants such as citrate, EDTA, or heparin), plasma, serum, erythrocyte sedimentation rate brown layer, lymphocytes, cell-free body fluids, and other samples. This product can process 0.1-1 mL of whole blood with a maximum yield of 30% μ g. It can purify DNA with sizes ranging from 100 bp to 50 kb. The purified DNA has high yield and good quality, and can remove protein, pigment, lipid, and other inhibitory impurities to the maximum extent. It can be directly used for PCR, fluorescence quantitative PCR, enzyme digestion, and Southern Blot experiments.
Self prepared reagent: anhydrous ethanol.
Preparation and important precautions before the experiment:
1. The sample should avoid repeated freeze-thaw cycles, otherwise it may result in smaller extracted DNA fragments and a decrease in extraction volume.
2. This reagent kit can extract up to 0.1-1 mL of whole blood samples or 1 × 107 white blood cells.
3.Before the first use, anhydrous ethanol should be added to Buffer GW1 and Buffer GW2 according to the instructions on the reagent bottle label.
4. Before use, please check if there is any crystallization or precipitation in the Buffer GL. If there is any crystallization or precipitation, please incubate the Buffer GL in a 56 ℃ water bath and dissolve it again.
5. The Buffer RCL in the reagent kit cannot be used again after being turbid.

Operation steps:
1. Sample processing: 1a When extracting 200 uL of blood sample, add the sample to the centrifuge tube (provided) and proceed directly to the next step of the experiment. 1b When the blood sample size is less than 200 μ When L, add Buffer GR to make up for 200 μ L. Proceed to the next step of the experiment. 1c When the blood sample size exceeds 200 μ When L is reached, add 1-2 times the volume of Buffer RCL, gently vortex or invert and mix well. Centrifuge at 12000 rpm (~13400 × g) for 1 minute and carefully discard the supernatant. If there is still red in the sediment, repeat the above steps once. Then add 200 to the precipitate μ Shake the buffer GR until thoroughly mixed before proceeding to the next step of the experiment. 1d If the processed blood sample is anticoagulant from poultry, birds, amphibians, or lower level organisms, its red blood cells are nucleated cells, and the blood sample size is 5-20 μ L. Can be added to Buffer GR to make up to 200 μ Follow up experiments will be conducted afterwards. Note: If downstream experiments are sensitive to RNA, 4 can be added μ L RNase A (100mg/mL) solution, shake for 15 seconds, and leave at room temperature for 5 minutes. RNase A reagent kit is not provided. If needed, you can order it separately from our company, item number: CW0601S.
2. Add 20 to the above solution μ L Protein K, mix well.

3. Add 200 μ Shake with L Buffer GL until thoroughly mixed. Note: Do not pre mix Protein K and Buffer GL.
4.Incubate at 4.56 ℃ for 10 minutes, invert and mix several times during this time. Attention: The DNA production has reached its maximum after 10 minutes of incubation, and further extension of incubation time has no effect on DNA production and purity.
5. Add 200 μ L anhydrous ethanol, invert and mix several times. Short centrifugation causes the liquid on the tube wall and wall cover to concentrate at the bottom of the tube.
6. Add all the solution obtained in step 5 to the spin columns DM that have been loaded into the collection tube. If the solution cannot be added at once, it can be transferred multiple times. Centrifuge at 12000 rpm for 1 minute, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube.
7. Add 500 to the adsorption column μ L Buffer GW1 (check if anhydrous ethanol is added before use), centrifuge at 12000 rpm for 1 minute, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube. Note: If the extracted sample is the blood genome of species such as mice or monkeys that are difficult to remove heme, it is recommended to repeat step 7.
8. Add 500 to the adsorption column μ L Buffer GW2 (check if anhydrous ethanol is added before use), centrifuge at 12000 rpm for 1 minute, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube. Note: To further improve DNA purity, repeat step 8.
9.Centrifuge at 9.12000 rpm for 2 minutes and discard the waste liquid from the collection tube. Place the adsorption column at room temperature for a few minutes to thoroughly air dry. Note: The purpose of this step is to remove residual ethanol from the adsorption column, which can affect subsequent enzymatic reactions (such as enzyme digestion, PCR, etc.)
10. Place the adsorption column in a new centrifuge tube (provided by oneself) and add 50-200 to the middle of the adsorption column in the air μ L Buffer GE or sterilized water, leave at room temperature for 2-5 minutes, centrifuge at 12000 rpm for 1 minute, collect DNA solution, and store DNA at -20 ℃. Note: 1) If downstream experiments are sensitive to pH or EDTA, they can be washed off with sterilized water. The pH value of the eluent has a significant impact on the elution efficiency. If water is used as the eluent, its pH value should be ensured to be between 7.0-8.5 (NaOH can be used to adjust the pH value of the water to this range). When the pH value is below 7.0, the elution efficiency is not high. 2) If the final concentration of DNA needs to be increased, the obtained DNA eluent can be added back to the adsorption membrane, left at room temperature for 2-5 minutes, and centrifuged at 12000 rpm for 1 minute. 3) Because DNA stored in water is affected by acidic hydrolysis, if long-term storage is required, it is recommended to elute with Buffer GE and store at -20 ℃.

安全和危险性（GHS）

SDS英文版

质量标准

质量标准

b665530_blood genomic dna mini kit.pdf

b665530_blood genomic dna mini kit..pdf

质检证书（CoA,COO,BSE/TSE 和分析图谱)

C of A & Other Certificates(BSE/TSE, COO):

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货号 (SKU)	包装规格	是否现货	价格	数量
B665530-50T	50T	期货
B665530-200T	200T	期货

血液基因组柱式小量提取试剂盒 (0.1-1 mL)

库存信息

库存信息

基本描述

安全和危险性（GHS）

质量标准

质检证书（CoA,COO,BSE/TSE 和分析图谱)

溶液计算器

摩尔浓度计算器

稀释计算器

复溶计算器