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Affymetrix/USB公司PrepEase® Genomic DNA Isolation Kit基因组DNA提取试剂盒评测
2012年06月02日 产品评测 评论数 2 ⁄ 被围观 6,220+


情况综述

产品

Affymetrix/USB公司PrepEase 基因组DNA提取试剂盒:PrepEase® Genomic DNA Isolation Kit

优点

快速纯化并得到基因组DNA

不用使用苯酚和柱子

在很短的时间内处理大量的样品

非常利于提取分子量较大的DNA

最大每次纯化可以达到 90 μg 产量

缺点

还是需要用氯仿来处理DNA,而且干燥的时间比较长

总结

一款物美价廉的DNA纯化试剂盒

For anyone that does a great deal of genotyping, the first step is always the isolation and purification of DNA from tissue or tail. I have tried several different kits and buffer systems in an attempt to find the purest, easiest, and least repetitive method of extraction. So when I heard about the USB PrepEase® Genomic DNA Isolation Kit, I thought I would give it a try.

The kit starts with the standard addition of buffer and proteinase K followed by a 55°C incubation. Although the kit claims that 2-4 hours is enough for complete DNA digestion, I have found that at least 6 hours is needed for digestion of hardier tail pieces, and even then there is always some skin and nail tissue left over. Following digestion is the addition of a protein precipitation buffer along with a 5:1 chloroform:isoamyl alcohol solution. This precipitates the protein from the solution, which is left in the lower aqueous phase after vortexing and spinning the sample at high speed for 4 minutes. The upper aqueous phase containing the DNA is collected, isopropanol is added to the solution and the DNA is precipitated out of solution to form a pellet. The pellet is subsequently subjected to an ethanol wash. The DNA pellet must then be dried for about an hour before rehydration with the proprietary DNA precipitation buffer. The yield is quite high, with a typical 30 ug per prep for a tail sample.

This kit works very well and the DNA isolated is pure, as I have never had any problems with downstream PCR reactions. The price per reaction is much less than Qiagen’s DNeasy Kit, which is my normal method of purification and the one I have found gives me the purest DNA. Although this kit gives results without the need for columns, it requires a smelly chloroform addition which requires the need for a fume hood and a lot of clean fresh air. I avoid working with chloroform if I can. That being my number one complaint, I have also never been fond of looking for tiny pellets on the bottom of tubes due to the possibility of a pellet being poured out along with the ethanol washes. There is also the wait time for the pellet to dry before reconstitution buffer can be added and the DNA used. Although the end result is just as good as any other kit, the process involves several steps that I would rather skip. If saving money is a main concern, it is also pretty easy to make your own digestion buffer and there are several protocols that use NaCl or similar compounds, eliminating the chloroform extraction process. This would be even cheaper than buying the USB kit. So, although, the DNA purification result is great, I don’t think I’ll be switching over. But if money is an issue and you’re happy spending a few extra minutes waiting for the pellet to dry and you don’t mind the chloroform odor, this would be a perfect fit.

产品信息

Part # Description Unit Size Your Price(USD)
Qty
78850 1 KT
PrepEase Genomic DNA Isolation Kit
50 preps Contact Distributor
78855 1 KT
PrepEase Genomic DNA Isolation Kit
250 preps Contact Distributor

产品相关资料下载:

MSDS / Material Safety Data Sheets

PrepEase Genomic DNA Isolation Kit_MSDS

Protocol, Long

PrepEase Genomic DNA Isolation Kit_Protocol



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