DNA Extraction from Agarose Gels
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FAQ
| Pergunta | Resposta |
|---|---|
| What recovery of my DNA can I expect with the Montage DNA Gel Extraction Kit? | Recovery is: >75 % for DNA less than 1kb >50% to <75% for DNA 1-2 kb <50% for DNA >2kb |
| What happens if the I increases the g-force with the Montage Gel Extraction Kit centrifugal device? | The nebulizer orifice is sized for gel extrusion at the documented g-force. The gel may clog the orifice at a higher g-force, which may affect DNA recovery. |
| Since Micropure is no longer available, what product should I use to do elution of DNA from agarose? | The Montage Gel Extraction Kit can now be used in place of Micropure for elution of DNA from agarose. |
| What is the pore size of the filter in the Montage Gel Extraction Kit? | The pore size is 0.45u. |
| Should I freeze my gel before adding it to the Montage Gel Extraction Kit? | The Montage Gel Extraction Kit allows for quick removal of DNA from Agarose without freezing. The only preparation required is to cut and trim the band of interest from the gel. The recovery of DNA is optimized by including the nebulizer, so there is no need for additional gel preparation. |
| The operating instructions for the Ultrafree DA recommend making up a modified TAE buffer. Why use modified TAE rather than standard TAE? | The modified TAE has 1/10th the final concentration of Na2EDTA of standard TAE. The activity and specificity of many enzymes used in molecular biology is magnesium-dependent. The lower level of EDTA ensures that the activity of enzymes that you may use down stream is not influenced by EDTA lowering the effective concentration of magnesium (i.e., if present, higher levels of EDTA could sequester much of the magnesium). The new Montage Gel Extraction Kit comes with 500mL of 50x stock modifies TAE buffer. |
| Can I add more than one band per device? | It is not recommended. When the amount of gel increases, the recovery decreases. |
| 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. |
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