BioMicroCenter:PippinPrep: Difference between revisions
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The BluePippin (Sage Science) is an automated preparative gel electrophoresis system that simplifies size fractionation of DNA. Runs of the five lane gels take approximately 1 hour and can either use internal markers for calibration or ethidium bromide to detect passing bands. The BluePippin can separate bands between 50bp and 50kb. We have done some testing of the BluePippin system and the results are [[BioMicroCenter:PippinPrep_Testing|'''HERE''']]. | The BluePippin (Sage Science) is an automated preparative gel electrophoresis system that simplifies size fractionation of DNA. Runs of the five lane gels take approximately 1 hour and can either use internal markers for calibration or ethidium bromide to detect passing bands. The BluePippin can separate bands between 50bp and 50kb. We have done some testing of the BluePippin system and the results are [[BioMicroCenter:PippinPrep_Testing|'''HERE''']].<BR><BR> | ||
[https://mit-ki.ilabsolutions.com/equipment/show/262716/?tab=schedule SCHEDULE TIME ON THE PIPPIN] | |||
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| [[Image:BMC_pippinGel.png|thumb|left|250px]] | | [[Image:BMC_pippinGel.png|thumb|left|250px]] |
Revision as of 09:26, 28 November 2014
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The BluePippin (Sage Science) is an automated preparative gel electrophoresis system that simplifies size fractionation of DNA. Runs of the five lane gels take approximately 1 hour and can either use internal markers for calibration or ethidium bromide to detect passing bands. The BluePippin can separate bands between 50bp and 50kb. We have done some testing of the BluePippin system and the results are HERE. |
The system is based on precast gel cartridges. These gels narrow as they run, concentrating the sample from the initial 50ul loaded in the wells. The BluePippin uses an optical detector to measure the rate of passage of bands based on high speed fluorescent markers or by directly detecting the DNA using ethidium bromide. Once the desired band passes the detector, the valve on each lane switches from a waste reservoir to an elution chamber. Once the band is passed, the valve turns back to waste and the desired band can be removed from the elution chamber with a pippette. Samples can be cleaned up with either SPRI beads or using an ethanol precipitation. |
GEL | SIZE RANGE | NOTES |
---|---|---|
3.0% | 50-200bp | Used for small RNA libraries |
2.0% | 100-600bp | |
1.5% | 0.3-1.5kb | Used for RNAseq libraries |
0.75% | 1.0-50kb | Uses pulse field. |
The BluePippin has four different percentages of agarose gels that can be used for different size selections. A stock of most types of cartridges are kept in the BioMicro Center. For long fragments using the pulse field, there are mutliple settings on the BluePippin, each optimized for a different size range. All 5 lanes of the gel should be used at once. Pricing for the cartridges and for training can be found on the |PRICING PAGE.