Extracción de ADN en geles de agarosa
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Preguntas frecuentes
| Pregunta | Respuesta |
|---|---|
| Can I use low melting point (LMP) Agarose with the the Montage Gel Extraction Kit? | Millipore has not tested the recoveries with LMP Agarose, but because this Agarose is so soft and deformable, the recoveries are not expected to be as high as recoveries observed when using the Montage Gel Extraction Kit with standard Agarose. |
| Can I use the Montage Gel Extraction Kit with low melt agarose ( LMP )? | No. LMP Agarose will not work in the Montage Gel Extraction Kit. However, for DNA fragments >125 base pairs the Centricon-100 is an excellent choice. |
| Can I use the Montage Gel Extraction Kit with DNA which is larger than 1000bp? | Yes, the Montage Gel Extraction Kit is recommended for extraction of DNA up to 10,000bp from agarose gel. For high recovery of larger DNA we recommend using the Centrilutor Micro-Electroelutor or using our protocol for recovery of DNA from LMP Agarose with Microcon-100. |
| Can I add more than one band per device? | It is not recommended. When the amount of gel increases, the recovery decreases. |
| Does the Ultrafree-DA come preassembled? | Yes. Although it consists of 3 subunits, it comes preassembled and ready to use. |
| Does the use of standard TAE interfere with the removal of DNA from the agarose gel? | No. You can still get removal of your DNA from the gel slice using the standard TAE. The additional EDTA in the buffer can cause problems downstream, as the EDTA can interfere with enzyme activity. If this is a problem, one can either use the modified TAE or do a buffer exchange in a Microcon to eliminate the EDTA. |
| How can I improve recovery when using the Montage Gel Extraction Kit? | Best recovery is achieved by working with <1.25% Agarose. Use a single band and trim off any excess gel. Spin at 5,000xG for approximately 10 minutes. |
| I'm using the Montage Gel Extraction Kit to extract DNA from agarose according to the operating instructions which recommend using TAE buffer. What is the advantage of TAE over TBE ? | Since you are extruding the DNA along with buffer from a gel slice, it is important that the electrophoresis buffer is compatible with downstream manipulations such as cloning and DNA sequencing. We found that TBE inhibits DNA sequencing. TAE, on the other hand, is a perfectly good electrophoreses buffer and all the enzymes that we have tested are fully functional in 1X TAE. |
| How do I order a gel nebulizer? | Gel nebulizers are a product enhancement included with the Montage Gel Extraction Kit. They increase the recovery of DNA by macerating the Agarose gel, thus facilitating DNA migration from the gel. |
| I wish to extract DNA from an agarose gel. Does the Montage Gel Extraction Kit also remove ethidium bromide? | The ethidium bromide stain and the DNA will pass through the Montage Gel Extraction Kit. For cloning and sequencing, there is no need for ethidium bromide removal after purification with the device. If you wish to remove the ethidium bromide, we recommend you take the DNA solution that has passed through the Montage Gel Extraction Kit and place it in the designated Microcon (based on the size of the DNA) and add 80% isopropanol. The alcohol solution will remove the ethidium bromide stain and the DNA will be concentrated in the Microcon. |
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