噬菌体 DNA 分离试剂盒

概述

该试剂盒提供了一种快速旋转柱法，可从在液体培养物中生长的细菌中繁殖的多种噬菌体中纯化总 DNA。DNA 的分离无需使用苯酚、氯仿或氯化铯带状程序。这种基于旋转柱的程序非常快速，可在 45 分钟内完成。该试剂盒可高效处理小体积噬菌体上清液（500 µL - 1 mL），通过可选的 DNase 和蛋白酶 K 处理，可最大限度地提高噬菌体 DNA 产量，同时将宿主 DNA 污染降至最低。纯化的总噬菌体 DNA 具有最高的完整性，可用于多种下游应用，包括 PCR、qPCR、限制性片段长度多态性 (RFLP)、测序、克隆、Southern 印迹等。

细节

Supporting Data

Figure 1 / 2

Figure 1. Effective Host Genomic DNA Removal without Reducing Phage DNA Yield.

Total DNA was isolated from four enriched phage cultures using Norgen's Phage DNA Isolation Kit. A DNase I pre-treatment was performed prior to adding the provided Lysis Buffer. Briefly, 20 units of DNase I was added to 1 mL of enriched phage culture and the mixture was incubated at room temperature for 20 minutes. After the DNAase I treatment the procedure was followed. As a control, DNA was isolated from aliquots of the same 4 cultures using Norgen’s Phage DNA Isolation Kit without performing the DNase I treatment. For DNA analysis 10 µL of each 50 µL elution was loaded onto a 1X TAE agarose gel. As it can be seen, the phage DNA was safely protected from the DNase I treatment by its coat protein, while the host genomic DNA was efficiently degraded by the DNase I. Thus the DNase I pre-treatment resulted in less host gDNA contamination in the final phage elution without influencing the total phage DNA yield. Lane M is Norgen's Highranger 1 kb DNA Ladder (Cat. 11900)

Optional Proteinase K Treatment Improves Yield — Figure 2. Optional Proteinase K Treatment Improves DNA Yield for Certain Phage Strains.

Total DNA was isolated with and without the optional Proteinase K treatment using Norgen's Phage DNA Isolation Kit. Briefly, 4 µL of Proteinase K (20 mg/mL) was added to 1 mL of enriched phage culture and incubated at 55°C for 15 minutes with the phage Lysis Buffer. After the Proteinase K treatment the procedure was followed. As a control, DNA was isolated from aliquots of the same 8 cultures using Norgen's Phage DNA Isolation Kit without performing the Proteinase K treatment. For DNA analysis 10 µL of each 50 µL elution was loaded onto a 1X TAE agarose gel and the yield of DNA was compared from the eight different phage types (lane 1 to 8). As it can be seen, the optional treatment of Proteinase K improved the phage DNA yield in Lanes 2, 5 and 6 dramatically. Lane M is Norgen's Highranger 1 kb DNA Ladder (Cat. 11900)

支持数据

图 1.有效去除宿主基因组 DNA，同时不降低噬菌体 DNA 产量使用 Norgen 的噬菌体 DNA 分离试剂盒从四个富集的噬菌体培养物中分离出总 DNA。在加入所提供的裂解缓冲液之前，先进行 DNase I 预处理。简而言之，在 1 mL 富集的噬菌体培养液中加入 20 个单位的 DNase I，然后将混合物在室温下培养 20 分钟。DNA 酶 I 处理后，按程序进行。作为对照，使用 Norgens 噬菌体 DNA 分离试剂盒从相同的 4 个培养物中分离出 DNA，但未进行 DNase I 处理。为了进行 DNA 分析，将每 50 µL 洗脱液中的 10 µL 加载到 1X TAE 琼脂糖凝胶上。由此可见，噬菌体 DNA 在其外壳蛋白的保护下安全地免受 DNase I 处理，而宿主基因组 DNA 则被 DNase I 有效降解。M 泳道为 Norgen's Highranger 1 kb DNA Ladder（类别号：11900)

图 2.可选的蛋白酶 K 处理可提高某些噬菌体菌株的 DNA 产量。使用 Norgen 的噬菌体 DNA 分离试剂盒分离了经蛋白酶 K 处理和未经蛋白酶 K 处理的总 DNA。简而言之，在 1 mL 富集噬菌体培养液中加入 4 µL 蛋白酶 K（20 mg/mL），然后与噬菌体裂解缓冲液一起在 55°C 孵育 15 分钟。蛋白酶 K 处理后，按照程序进行。作为对照，使用 Norgen 的噬菌体 DNA 分离试剂盒从相同的 8 个培养物的等分中分离出 DNA，但未进行蛋白酶 K 处理。为了进行 DNA 分析，将每 50 µL 洗脱液中的 10 µL 加载到 1X TAE 琼脂糖凝胶上，比较八种不同类型噬菌体（第 1 至第 8 泳道）的 DNA 产量。可以看出，蛋白酶 K 的可选处理大大提高了 2、5 和 6 泳道中噬菌体 DNA 的产量。M 泳道为 Norgen's Highranger 1 kb DNA Ladder（类别号：11900)

试剂盒规格
色谱柱结合能力	50 µg
最大色谱柱装载量	650 µL
纯化的 DNA 大小	所有大小
起始材料的最大用量	1 x 10¹⁰ pfu/mL 富集噬菌体
平均产量*	3-15 µg de ADN a partir de 10⁶-10¹⁰ pfu/mL 丰富的噬菌体
完成 10 次净化所需时间	45 分钟

* 平均产量因使用的条件和发育阶段而异。

储存条件和产品稳定性
所有溶液都应密封保存在室温下。该试剂盒在发货后 1 年内保持稳定。

Alternate formats available upon request by emailing info@norgenbiotek.com. Such requests shall be made:

in a timely manner that considers the person's accessibility needs due to disability; and
at a cost that is no more than the regular cost charged to other persons

概述

细节

Supporting Data

Figure 1. Effective Host Genomic DNA Removal without Reducing Phage DNA Yield.

Figure 2. Optional Proteinase K Treatment Improves DNA Yield for Certain Phage Strains.

支持数据

Documentation

PROTOCOLS

SAFETY DATA SHEETS

FLYERS

Your Shopping Cart is Empty