As we have only a UV transilluminator, i am afraid to damage my DNA while viewing with UV light, so can i purify it with PCR purification kit rather than run an agarose gel and purify it.
P.S: i am using BamHI, which non thermo sensitive.
1. As long as you do it reasonably quick, your DNA should be fine from UV transilluminator. Many researchers worldwide have done gel-extraction procedures without obvious problems.
2. If you want to use a kit, you need to read the instruction of the kit very carefully. The requirements for using a specific kit can be vary. For example, Qiagene sells these two products. One is QIAquick PCR Purification Kit (as you mentioned) for PCR product purification. And the other one is designed for cleaning up restriction digestion products, including removing restriction enzymes, called MinElute Reaction Cleanup Kit ( https://www.qiagen.com/us/shop/sample-technologies/dna/dna-preparation/minelute-reaction-cleanup-kit/#productdetails ).
The optimum size ranges of DNA recovered from these two kits are different. So, you need to be careful.
(a) PCR purification kit: 100 bp to 10 kb
(b) MinElute Reaction Cleanup kit: purification of DNA 70 bp – 4 kb in size from enzymatic reactions
Hello Oualid,
If the restriction results in linearizion of a plasmid vector than of-course there is no need to cut it out from the gel and it can be directly purified using a PCR cleanup kit. However, if restriction results in two fragments then you have to excise out the desired fragment from the gel. There are few ways to minimize the DNA nicking resulting from UV exposure. Use long-wavelength UV while minimizing the time of exposure as suggested by Yuan-Yeu. Another measure is to put the gel on a thick glass (or less preferably transparent plastic) plate. This will not block but minimize the UV penetration. Best option is of-course to use a blue light trans-illuminator. I purchased one from the link given below. This blue light trans-illuminator is perhaps the cheapest (US$ 80) transilluminator and I am a very happy user of it.
http://iorodeo.com/products/mini-led-transilluminator
Thank you very much for your answers.
Yuan, effectively that's what i am suspecting, the kit its not suitable for restriction reaction.GeneAll have a kit for general DNA cleaning but not Qiagen.
Syed Ali thanks to you as well, i want also to purchase a trans illuminator, your feedback is very useful to me.
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