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Discover Bio

 Manual Microwave Peptide Synthesizer

600423 • Revision 2 • August 2017

9

Recommended Synthesis Parameters

Recommended Microwave Methods

Recommended Reagents for 0.1 mmol Scale

Activator Base Solution

Prepare a solution of ~0.3 M DIEA in DMF.

Deprotection Solution

Prepare a solution of 20% piperidine in DMF with 0.1 M HOBt monohydrate.

Method

Power

Max Temp.

Run Time

Delta Temp.

Deprotection

20 W

75 °C

3:00

5 °C

Coupling

20 W

75 °C

5:00

5 °C

TFA Cleavage

15 W

38°C

30:00

5 °C

NOTE

Cysteine and histidine couplings should be performed at 50 °C to reduce racemization.  Cleavage 

temperatures should never exceed 43 °C.  All values for Power are suggested, and should be 

adjusted depending on reaction scale.  The goal is to reach the maximum temperature in 75 

seconds for Deprotection and Coupling, and in 120 seconds for Cleavage.

Number of 

Couplings

0.1 mmol Scale

mL 

DMF

mL 

DIEA

Final 

Vol.

5

19

1

20

10

38

2

40

Per Coupling:

4 mL

Final Volume

0.1 mmol Scale

mL 

DMF

mL 

Pip.

g HOBt 

monohydrate

50

40

10

0.77

100

80

20

1.5

500

400

100

7.7

Per Coupling:

7 mL

NOTE

HOBt is optional, but is recommended for sequences susceptible to aspartimide formation.

WARNING

The Discover Bio reaction vessels should not be operated above 75 

º

C.

Содержание Discover Bio

Страница 1: ...g Method 8 Recommended Synthesis Parameters 9 Recommended Microwave Methods 9 Recommended Reagents for 0 1 mmol Scale 9 General Discover Bio Methods 11 Swelling the Resin 11 Performing a Deprotection 12 Performing a Coupling 12 Performing a Cleavage 13 Using the 4 mL Vessel for Smaller Scale 15 Connecting the 4 mL Vessel 15 Maintenance Procedures 16 After Each Use 16 Monthly Maintenance 16 Semiann...

Страница 2: ...peptide synthesis system Built on CEM s flexible Discover microwave platform the Discover Bio allows the rapid and efficient synthesis of peptides on scales from 0 05 to 0 5 mmol thanks to the efficient washing station and CEM s patented circular microwave cavity Product Position Nitrogen Valve Left Knob Add Port Right Knob Power Switch Discover Bio Microwave Reactor Discover Bio Wash Station Reag...

Страница 3: ...e Discover Bio requires electrical power of 120 VAC 60 Hz 1 7 A or 240 VAC 50 Hz 1 7 A where applicable Specific power requirements 120 VAC vs 240 VAC can be found on the nameplate affixed to the rear of the Discover Bio Microwave Reactor and on the rear of the Discover Bio Wash Station Safety Information Fume Ventilation The Discover Bio operates as a semi closed system with minimal venting of an...

Страница 4: ... the bottle tubing to the rear of the Wash Station The tube labeled LIQ should be connected to position 4 The unlabeled tube should be connected to position 1 3 Set up the Deprotection bottle 3 1 Connect a bottle cap to the 1 L bottle to be used for deprotection solution Ensure the dip tube reaches the bottom of the bottle and that the dip tube filter is in place 3 2 Connect the bottle tubing to t...

Страница 5: ...ubing should be run into a ventilated fume hood Product Collection Position Setup 1 Cut two pieces of tubing approximately 18 45 cm 2 Take one piece of tubing and using yellow ferrules and nuts connect one end to the left port on the rear of the Product Collection Position then connect the other end to position 10 3 Take one piece of tubing and using yellow ferrules and nuts connect one end to the...

Страница 6: ...well is just resting on the bottom of the vessel Secure the thermowell with the nut 4 Connect the reaction vessel vent line 4 1 Cut a length of tubing 48 120 cm Using a yellow ferrule and nut connect the tubing to the other side port of the reaction vessel top being careful to bend the sprayhead line out of the way 4 2 Using a yellow ferrule and nut connect the other end of the tubing to position ...

Страница 7: ...priate power source Locate the power switch on the left side of the Discover and turn the power on 2 After the Discover has started up press the EDIT key to open the System Menus 3 The first menu option will be Temperature Press ENTER to open the Temperature Menu 4 Use the right arrow key to scroll to Select Alternate and press ENTER 5 Use the right arrow key to change to DEVICE FIBER OPTIC and pr...

Страница 8: ...r setting in W and press ENTER 5 Using the numeric keypad enter the maximum temperature setting in C and press ENTER 6 Using the numeric keypad enter the run time in min sec and press ENTER 7 Using the numeric keypad set the Delta Temperature value to 5 C and press ENTER 8 Using the right arrow key set stirring to OFF and press ENTER 9 Using the right arrow key set cooling to OFF and press ENTER 1...

Страница 9: ... couplings should be performed at 50 C to reduce racemization Cleavage temperatures should never exceed 43 C All values for Power are suggested and should be adjusted depending on reaction scale The goal is to reach the maximum temperature in 75 seconds for Deprotection and Coupling and in 120 seconds for Cleavage Number of Couplings 0 1 mmol Scale mL DMF mL DIEA Final Vol 5 19 1 20 10 38 2 40 Per...

Страница 10: ... 0 190 Fmoc Asp OtBu OH 411 5 0 206 0 190 Fmoc Cyc Trt OH 585 7 0 293 0 190 Fmoc Gln Trt OH 610 7 0 305 0 190 Fmoc Glu OtBu OH 425 5 0 213 0 190 Fmoc Gly OH 297 3 0 149 0 190 Fmoc His Trt OH 619 7 0 310 0 190 Fmoc Ile OH 353 4 0 177 0 190 Fmoc Leu OH 353 4 0 177 0 190 Fmoc Lys Boc OH 468 5 0 234 0 190 Fmoc Met OH 371 5 0 186 0 190 Fmoc Phe OH 387 4 0 194 0 190 Fmoc Pro OH 337 4 0 169 0 190 Fmoc Se...

Страница 11: ... 3 Slide the drain line out of the groove in the top of the vessel assembly Open the reaction vessel by pressing the vessel body up and rotating Add the resin to the vessel Seal the vessel by pressing the vessel body up into the vessel top and rotating Slide the drain line back into the groove in the top of the vessel assembly Place the vessel in the reaction vessel holder 4 Set the left knob to W...

Страница 12: ...sure the fiber optic probe is fully inserted into the thermowell 5 Insert the reaction vessel into the Discover cavity Load the Deprotection microwave method and press the Play Pause key to begin the method 6 When the microwave method is finished remove the reaction vessel from the Discover cavity and place it in the vessel holder Set the left knob to N2 Purge Wash the right knob to Waste 7 When a...

Страница 13: ... the left knob to Wash Solvent and add approximately 7 mL of solvent to the vessel then set the left knob to Stop Optionally to bubble the resin for a more efficient wash turn the right knob to N2 Bubbling and allow the resin to bubble for five seconds 11 Set the left knob to N2 Purge Wash then set the right knob to Waste to drain the reaction vessel Set both knobs to Stop 12 Repeat steps 10 and 1...

Страница 14: ... Discover cavity and place it in the vessel holder 11 Connect a clean labeled centrifuge tube to the product manifold 12 Turn the right knob to Product to drain the cleaved peptide into the centrifuge tube 13 Remove the centrifuge tube containing the cleaved peptide The peptide is now ready to be precipitated and analyzed 14 Set the right knob to Stop Set the left knob to Wash Solvent and add appr...

Страница 15: ...es must be added manually by opening the top of the reaction vessel Connecting the 4 mL Vessel 1 Place a yellow ferrule and nut on the end of the thermowell Insert the thermowell into the center port of the reaction vessel top so that the bottom of the thermowell is on the bottom of the vessel Secure the thermowell with the nut 2 Connect the RV Vent line to the side port of the reaction vessel top...

Страница 16: ...With the pump turned on turn the right knob to Product to drain the dichloromethane into the centrifuge tube Repeat with an additional 20 mL dichloromethane Monthly Maintenance Replace the dip tube filters in the main solvent and deprotection bottles Semiannual Maintenance Replace the Wash Solvent with methanol and perform a wash of the reaction vessel Replace the frit on the bottom of the reactio...

Страница 17: ...4 Using the frit removal tool pop the frit out of the vessel body 5 Insert a new frit into the vessel body 6 Screw the vessel body into the bottom of the reaction vessel Close the reaction vessel by pressing up into the reaction vessel top and turning counterclockwise Connect the drain line to the bottom of the reaction vessel ...

Страница 18: ...ply 908400 240 v 50 Hz Power Supply 908405 Power Cord varies by country Fiber Optic Temperature Control 541176 Discover Bio Accessory Kit 908410 Product Part Number 4 mL Reaction Vessel Assembly 543505 Replacement Frits for 4 mL Vessel Qty 50 215005 25 mL Reaction Vessel Assembly 543500 Replacement Frits for 25 mL Vessel Qty 50 215055 50 mL Polypropylene Centrifuge Tubes Qty 25 167150 M 1 L Glass ...

Страница 19: ...lown fuses Replace consumables and Clean the instrument as required How to Get Service Purchaser should contact the Service Department of CEM Corporation or the nearest CEM subsidiary or dis tributor for return authorization and for proper crating and shipping instructions to return instrument freight prepaid for service On site repairs by an authorized service technician are available through the...

Страница 20: ... Business Park Damastown Dublin 15 Tel 353 0 1 885 1752 Fax 353 0 1 885 1601 Email info ireland cem com www cemmicrowave co uk Italy CEM S R L Via Dell Artigianato 6 8 24055 Cologno al Serio Bg Tel 39 35 896224 Fax 39 35 891661 Email info srl cem com www cemmicroonde com Japan CEM Japan K K 2 18 10 Takanawa Minato ku Tokyo 108 0074 Tel 81 3 5793 8542 Fax 81 3 5793 8543 Email info cemjapan jp www c...

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