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Summary of Contents for Biotage PhyPrep
- Page 1 Biotage PhyPrep ® User Manual...
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Page 2: Table Of Contents
Gigaprep 1 Sample Deck Layout Gigaprep 2 Samples Method Gigaprep 2 Samples Deck Layout Gigaprep 3 Samples Method Gigaprep 3 Samples Deck Layout Gigaprep 4 Samples Method Gigaprep 4 Samples Deck Layout Biotage PhyPrep User Manual | © Biotage 2021 ®... - Page 3 Biotage® PhyPrep...
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Page 4: System Overview
System Overview System Overview The Biotage PhyPrep system is the only all-in-one instrument for automated endotoxin-free plasmid purification in maxi, mega, and ® giga scales. This system offers a variety of user friendly protocols for endotoxin-free plasmid purification. It is developed and supported by scientists and engineers to provide consistent high-quality endotoxin-free plasmids for transient transfection. -
Page 5: Hardware & Connections
7. Waste port and blue tubing Panel PC Vacuum pump Leakage Tray Vacuum Vacuum Waste Carboy Waste Carboy Parts Vacuum Waste Sample Elution Tube Wash Buffer Vacuum Växel Transfer Elution PhyPrep Carboy Carboy Filter Stand Reservoir Column Tube Tube Reservoir... -
Page 6: Instrument Safety
Sample Filter Reservoir Emergency Stop Assembly The PhyPrep door works as an emergency stop. If the door is Assemble the Sample Filter Reservoirs with the correct orientation opened at any time during system operation, the instrument before placing them on the deck. The reservoir consists of a filter will stop operating immediately, and the current method will be top and a reservoir bottom. -
Page 7: Biotage ® Phyprep Plasmid Kit Endotoxin-Free
Biotage® PhyPrep Plasmid Kit Endotoxin-Free Biotage PhyPrep Plasmid Kit Endotoxin-Free ® Kit Contents Maxiprep kit DPM-16-05-72-KIT DPM-16-05- DPM-00- 710-0001 900-0078 900-0081 900-0084 900-0091 900-0093 900-0087 900-0089 700-0030 417458SP 72-RNA 25-00 16 x 16 x 16 x 16 x 16 x... -
Page 8: Instrument Deck
Tube Figure 1. The PhyPrep deck layout 1) Transfer tubes 2) Vacuum station 3) Wash Buffer Reservoir 4) Biotage Växel columns and PhyPrep Tubes 5) Supplemental buffers 6) Biotage Växel columns and PhyPrep Tubes 7) Elution Tube Stand 8) Sample Filter Reservoirs Biotage PhyPrep User Manual | ©... -
Page 9: Before You Begin
600–750 mL of an overnight bacterial culture. » Centrifuge at ≥ 3500 x g for 30 minutes. Discard supernatant. » The PhyPrep deck can be set up while waiting for cells to pellet. See appropriate deck layout per desired method for further instructions. -
Page 10: Sample Preparation Guide
» Place Sample Filter Reservoir on the PhyPrep deck. Then slowly pour entire contents of the centrifuge bottle into the middle of the Sample Filter Reservoir to allow the supernatant to go through the filter first. This will prevent the precipitated particulates clogging the filter. - Page 11 Sample Preparation Guide Sample Preparation Guide Required Kit RNase A Resuspension Buffer Lysis Buffer Precipitation Buffer Thawed Cell Pellet ADD TO THEN ADD TO Shake until pellet is 1. Resuspend Cells completely resuspended (20-30 seconds) THEN RNase A Resuspension Thawed or Fresh Buffer Cell Pellet ADD TO...
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Page 12: Maxiprep 1 Sample Method
Uncap and place 1 x 50 mL pyrogen-free conical tubes into position A. » Elution collection tube should rest in the bottom portion of the elution stand. » Place the Elution Tube Stand back onto the PhyPrep deck. Section » Place 1 x Sample Filter Reservoir into A. -
Page 13: Maxiprep 1 Sample Deck Layout
Elution Elution Tube Tube Sample Sample Tube Tube Tube Tube Sample Sample Sample 1 Sample 2 Tube Tube Sample Sample Tube Tube Elution Elution PhyPrep Tube Tube Tube Sample 3 Sample 4 Sample 1 Sample 2 Sample 3 Sample 4... -
Page 14: Maxiprep 2 Samples Method
Uncap and place 2 x 50 mL pyrogen-free conical tubes into positions A and B. » Elution collection tubes should rest in the bottom portion of the elution stand. » Place the Elution Tube Stand back onto the PhyPrep deck. Section » Place 2 x Sample Filter Reservoirs into A and B. -
Page 15: Maxiprep 2 Samples Deck Layout
Tube Tube Sample Sample Tube Tube Tube Tube Sample Sample Sample 1 Sample 2 Tube Tube Sample Sample Tube Tube Elution Elution PhyPrep PhyPrep Tube Tube Tube Tube Sample 3 Sample 4 Sample 1 Sample 2 Sample 3 Sample 4... -
Page 16: Maxiprep 3 Samples Method
Uncap and place 3 x 50 mL pyrogen-free conical tubes in position A, B, and C. » Elution collection tubes should rest in the bottom portion of the elution stand. » Place the Elution Tube Stand back onto the PhyPrep deck. Section » Place 3 x Sample Filter Reservoirs into A, B, and C. -
Page 17: Maxiprep 3 Samples Deck Layout
Elution Tube Tube Sample Sample Tube Tube Tube Tube Sample Sample Sample 1 Sample 2 Tube Tube Sample PhyPrep Tube Elution Elution Tube PhyPrep PhyPrep Tube Tube Tube Tube Sample 3 Sample 4 Sample 1 Sample 2 Sample 3 Sample 4... -
Page 18: Maxiprep 4 Samples Method
Uncap and place 4 x ELUTION BUFFER tubes into A, B, C and D. » Place 2 x PhyPrep Tubes into E and F. Add 40 mL of ENDOTOXIN REMOVAL BUFFER to these tubes. » Place 2 x PhyPrep Tubes into A and B. -
Page 19: Maxiprep 4 Samples Deck Layout
Elution Elution Tube Tube Sample Sample Tube Tube Tube Tube Sample Sample Sample 1 Sample 2 Tube Tube PhyPrep PhyPrep Elution Elution Tube Tube PhyPrep PhyPrep Tube Tube Tube Tube Sample 3 Sample 4 Sample 1 Sample 2 Sample 3... -
Page 20: Megaprep 1 Sample Method
Add 270 mL of WASH BUFFER to the Wash Buffer Reservoir. » Place 3 x PhyPrep Tubes in A, C, and E. Section » Place 1 x PhyPrep Tube into I. Add 30 mL of EQUILIBRATION BUFFER to this tube. -
Page 21: Megaprep 1 Sample Deck Layout
Elution Elution Buffer Buffer Buffer Buffer Sample Sample Tube Tube Sample Sample Tube Tube Sample Sample Tube Tube Sample PhyPrep Elution Elution Tube Tube Sample Sample Tube Tube Tube Tube Sample PhyPrep Sample 1 Sample 2 Tube Tube Sample Sample... -
Page 22: Megaprep 2 Samples Method
Add 470 mL of WASH BUFFER to the Wash Buffer Reservoir. » Place 6 x PhyPrep Tubes into A, B, C, D, E, and F. Section » Place 2 x PhyPrep Tubes into I and J. Add 30 mL of EQUILIBRATION BUFFER to these tubes. -
Page 23: Megaprep 2 Samples Deck Layout
Buffer Elution Elution Buffer Buffer Buffer Buffer Sample Sample Tube Tube Sample Sample Tube Tube Sample Sample Tube Tube PhyPrep PhyPrep Elution Elution Tube Tube Sample Sample Tube Tube Tube Tube PhyPrep PhyPrep Sample 1 Sample 2 Tube Tube Sample... -
Page 24: Megaprep 3 Samples Method
Add 670 mL of WASH BUFFER to the Wash Buffer Reservoir. » Place 6 x PhyPrep Tubes into A, B, C, D, E, and F. Section » Place 2 x PhyPrep Tubes into I and J. Add 30 mL of EQUILIBRATION BUFFER to these tubes. -
Page 25: Megaprep 3 Samples Deck Layout
Växel Växel Buffer Buffer column column 30 mL 30 mL 30 mL Buffer Elution Elution Buffer Buffer Buffer Buffer PhyPrep Tube PhyPrep PhyPrep Elution Elution Tube Tube PhyPrep Tube Tube Tube PhyPrep PhyPrep Sample 1 Sample 2 Tube Tube... -
Page 26: Megaprep 4 Samples Method
Add 870 mL of WASH BUFFER to the Wash Buffer Reservoir. » Place 6 x PhyPrep Tubes into A, B, C, D, E, and F. Section » Place 2 x PhyPrep Tubes into I and J. Add 30 mL of EQUILIBRATION BUFFER to these tubes. -
Page 27: Megaprep 4 Samples Deck Layout
30 mL 30 mL Buffer Buffer Elution Elution Buffer Buffer Buffer Buffer Sample Sample Tube Tube Sample Sample Tube Tube PhyPrep PhyPrep Tube Tube PhyPrep PhyPrep Elution Elution Tube Tube PhyPrep PhyPrep Tube Tube Tube Tube PhyPrep PhyPrep Sample 1... -
Page 28: Gigaprep 1 Sample Method
Add 480 mL of WASH BUFFER to the Wash Buffer Reservoir. » Place 4 x PhyPrep Tubes in A, C, E, and G. Section » Place 1 x PhyPrep Tube into I. Add 40 mL of EQUILIBRATION BUFFER to this tube. -
Page 29: Gigaprep 1 Sample Deck Layout
40 mL 40 mL 40 mL 40 mL Buffer Buffer Elution Elution Buffer Buffer Buffer Buffer Sample Sample Tube Tube Sample PhyPrep Tube Tube Sample Sample Tube Tube Sample PhyPrep Elution Elution Tube Tube Sample Sample Tube Tube Tube... -
Page 30: Gigaprep 2 Samples Method
Add 880 mL of WASH BUFFER to the Wash Buffer Reservoir. » Place 8 x PhyPrep Tubes into A, B, C, D, E, F, G, and H. Section » Place 2 x PhyPrep Tubes into I and J. Add 40 mL of EQUILIBRATION BUFFER to these tubes. -
Page 31: Gigaprep 2 Samples Deck Layout
40 mL 40 mL 40 mL 40 mL Buffer Buffer Elution Elution Buffer Buffer Buffer Buffer Sample Sample Tube Tube PhyPrep PhyPrep Tube Tube Sample Sample Tube Tube PhyPrep PhyPrep Elution Elution Tube Tube Sample Sample Tube Tube... -
Page 32: Gigaprep 3 Samples Method
Add 1280 mL of WASH BUFFER to the Wash Buffer Reservoir. » Place 8 x PhyPrep Tubes into A, B, C, D, E, F, G, and H. Section » Place 2 x PhyPrep Tubes into I and J. Add 40 mL of EQUILIBRATION BUFFER to these tubes. -
Page 33: Gigaprep 3 Samples Deck Layout
Elution column Växel Växel Buffer Buffer column column 40 mL 40 mL 40 mL Buffer Elution Elution Buffer Buffer Buffer Buffer PhyPrep Tube PhyPrep PhyPrep Tube Tube PhyPrep Tube PhyPrep PhyPrep Elution Elution Tube Tube PhyPrep Tube Tube Tube PhyPrep... -
Page 34: Gigaprep 4 Samples Method
Add 1680 mL of WASH BUFFER to the Wash Buffer Reservoir. » Place 8 x PhyPrep tubes into A, B, C, D, E, F, G, and H. Section » Place 2 x PhyPrep Tubes into I and J. Add 40 mL of EQUILIBRATION BUFFER to these tubes. -
Page 35: Gigaprep 4 Samples Deck Layout
Växel Växel Buffer Buffer column column 40 mL 40 mL 40 mL 40 mL Buffer Buffer Elution Elution Buffer Buffer Buffer Buffer PhyPrep PhyPrep Tube Tube PhyPrep PhyPrep Tube Tube PhyPrep PhyPrep Tube Tube PhyPrep PhyPrep Elution Elution Tube Tube... -
Page 36: Software Overview
Check the boxes to confirm and press START. Select the number of samples. Then press NEXT. 6. Once the method is completed, press QUIT. Biotage PhyPrep User Manual | © Biotage 2021 ®... -
Page 37: Recommended Alcohol Precipitation Protocol
Recommended Alcohol Precipitation Protocols Recommended Alcohol Precipitation Protocols The elution buffer is 50 mM Tris-HCL, 500 mM NaCl, pH 8.5. 10. Carefully decant the ethyl alcohol and remove as much as If downstream assays require desalting, we recommend possible. the following alcohol precipitations. Note: Do not disturb the pellet. -
Page 38: Maintenance
Wash Buffer ejector pads buffer into it. Reservoir Biotage recommends a yearly Preventative Maintenance to be performed by a Biotage field engineer. Consult your Biotage representative for more information. Biotage PhyPrep User Manual | © Biotage 2021 ®... -
Page 39: Carboys Set Up And Care
Maintenance Carboys Set Up and Care A 20 L vacuum carboy and a 20 L waste carboy are included with the PhyPrep instrument. It is essential to empty the carboys once they reach 80% capacity. Prep Type 20 L Waste Carboy... -
Page 40: Troubleshooting
This troubleshooting guide is designed to help solve most problems that may arise while using the Biotage PhyPrep. The scientists at Biotage are always happy to answer any questions you may have about the information or protocols in this manual.For questions not addressed here, please contact Biotage support to talk with a Biotage scientist. -
Page 41: Low Dna Quality
Troubleshooting Low DNA quality Cause Improper sample preparation Explanations and actions » Incomplete precipitation . » Incomplete precipitation generates poor quality supernatant. Ensure that precipitation is complete by inverting the sample 30 times. Do not vigorously mix, this may cause additional contaminant release from the flocculent. -
Page 42: General Information
100 x 20 mL transfer tips for Biotage PhyPrep ® Biotage Växel columns (Ion Exchange) ® DPM-00-10-01 Sterile 100 x 20 mL transfer tips for Biotage ® Resuspension Buffer (60 mL bottles) PhyPrep Lysis Buffer (60 mL bottles) DPM-00-20-00 50 mL conical tubes with caps, sterile,... - Page 43 Notes Notes...
- Page 44 Notes Biotage PhyPrep User Manual | © Biotage 2021 ®...
- Page 45 Notes...
- Page 46 Literature Number: 417343-A © 2021 Biotage. All rights reserved. No material may be reproduced or published without the written permission of Biotage. Information in this document is subject to change without notice and does not represent any commitment from Biotage. E&OE.
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