MEGA-spin™ 凝胶DNA回收试剂盒_详细资料

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本试剂盒是既可从琼脂糖凝胶中，又可从PCR反应或其它各种酶促反应液中纯化单链或双链DNA片段的试剂盒

本试剂盒是既可从琼脂糖凝胶中，又可从PCR反应或其它各种酶促反应液中纯化单链或双链DNA片段的试剂盒，可有效除去酶蛋白、寡聚核苷酸引物、盐离子、dNTP等。吸附柱内采用独特的吸附材料，与常用硅胶膜相比，可更加GX、专一地吸附核酸分子。

使用本试剂盒操作简单方便，无需使用NaAC调节pH值，每次可纯化多至10μg的DNA片段（≥100bp)，纯化得率高达60～90%，可用20μl洗脱缓冲液进行洗脱，纯化得到的DNA片段可直接用于连接反应、PCR扩增、DNA测序等多种分子生物学实验。

主要特点：
1、独特的吸附材料：可更加GX、专一地吸附核酸分子；
2、纯化得率高：高达60～90%，可纯化10μg的DNA片段（≥100bp)；
3、纯度高：1.8＜A260/A280＜2.0，保证后续实验结果的可靠性和重复性；
4、操作简便、快速，只需15min即可完成全部操作；
5、使用安全：无须使用苯酚、氯仿等有毒试剂；
6、一盒两用：既可从PCR反应或其它各种酶促反应液，又可从中琼脂糖凝胶中纯化单链或双链DNA片段。

主要应用：
既可从PCR反应或其它各种酶促反应液，又可从琼脂糖凝胶中纯化单链或双链DNA片段，适用于一般常规分子生物学实验。

存储条件：
本试剂盒在室温（15-25℃）条件下可保存一年。

DNA fragments for probe DNA or ligation must be separatedpurified from other DNA fragments. MEGA-spin^TM employs a column method to purify target DNA in excised agarose gel. The column method uses a highly concentrated salt solution to keep the target DNA bound to the column membrane. The binding reaction occurs due to the disruption of the organized structure of water moleculesthe interaction with the nucleic acids. Thus the adsorption to the specifically pretreated membrane is favored. Since the binding process is specific for nucleic acids, the bound material can be separatedpurified from impurities e.g. saltsproteins, with simple washing step. Nucleic acids elute from the column membrane in a low salt buffer or water.

MEGA-spin^TM is designed to extractpurify DNA of 100bp to 10kb from standard or low-melt agarose gels in TAE or TBE buffer. Furthermore, the kit guarantees a high yield of purification up to 70-90%. DNA fragments isolated with MEGA-spin^TM Agarose Gel Extraction Kit are efficiently ligated into plasmid cloning vectors or specifically labeled using either random primed labeling or nick translation. No inhibition of digestion with restriction endonucleases is observed.

- Extractpurify DNA of 100bp to 10kb from all types of agarose gel
- Recovery between 7090%
- Spin column technology to recover DNA from excised agarose bands

- Sequencing, cloning, ligation, probe labeling ligation, random primed labeling, nick translationetc.

＊ Agarose Lysis Buffer
180㎖

＊ Washing Buffer
: Add Absolute EtOH 200ml before use.

50㎖

＊ Elution Buffer
20㎖

＊ Columns
: Column contains silica membrane

250 columns

＊ Collection Tubes

250 Tubes

1. % Recovery of DNA from the different fragment length & amount of DNA

[ Table. the different fragment length ] [ Table. the amount of DNA ]

Fragment length

% Recovery

100bp - 200bp

80%

200bp - 4Kb
90%

4Kb - 10Kb
85%

> 10 Kb
60%

Amount of DNA

% Recovery

5 - 10ug
90%

10 - 30ug
85%

> 30ug
70%

2. Recovery of DNA from different PCR product size

Purification of DNA fragments with the MEGA-spin^TM Agarose Gel Extraction Kit is based on solubilization of agaroseselective adsorption of nucleic acids onto MEGA-spin^TM silica-membrane in the presence of charotropic salt. The different sizes of DNA fragments can be extracted from gels in 70%-90% recovery.

Fig. DNA fragments extraction with the MEGA-spin^TM Agarose Gel DNA Extraction Kit
DNA fragments were extracted from gelspooled after extraction. Recoveries of approximately more than 70%-90% are obtained for different sizes (100bp-10kb).
Lane M1, M2, Marker DNA; Lane 1, 6, 131bp DNA; Lane 2, 7, 570bp DNA; Lane 3, 8, 1.3Kb DNA; Lane 4, 9, 4.5Kb DNA; Lane 5,10, 10Kb DNA

1. Enzyme digestion result

Fig. Enzyme digestion
3kb PCR product was extracted using MEGA-spin^TM Agarose Gel Extraction Kitthen digested with EcoRI. The digestions were Performed at 37¡E for 1hr with each enzymes. And then result was analyzedin gel electrophoresis.
Lane M, Marker DNA; Lane 1, 3, undigested PCR product, extracted using MEGA-spin^TM Kit; Lane 2,4, digested PCR product

The MEGA-spin^TM Kit was compared with other supplier''s products in the recovery of DNA fragments from gels. Comparison of various products at same conditions showed that the recovery of iNtRON''s MEGA-spin^TM Kit was similar or excellent to other products.

Fig. Comparison of different supplier''s products
The DNA fragments (4.5Kb1.3Kb) were extracted from gels using kits from different suppliers. 1.3Kb DNA fragment; 4.5 Kb DNA fragment
Lane M, Marker DNA; Lane 1, MEGA-spin^TM Kit; Lane 2, Supplier A; Lane 3, Supplier B;
Lane 4, Supplier C; Lane 5, Supplier D

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