Mini-PROTEAN® 3 Multi-Casting Chamber Instruction Manual
Catalog Number
165-4110
T able of Contents
Page Section 1 General Information (1)
雪中情1.1 Introduction (1)吴越版西游记
1.2 Specifications (1)
Section 2 Loading the Chamber (1)
Section 3 Casting Gels (2)
普通话证书有效期3.1 Casting Non-Gradient Gels from the Top (2)
3.2 Casting Linear and Convex Gradient Gels from the Bottom (3)
Section 4 Chamber Disasmbly and Storage of Gels (3)
Section 5 Using the Gels (4)
Section 6 Ordering Information (4)
Section 7 Reagents for Electrophoresis (5)
Appendix A Estimated Volume of Acrylamide for 12 Mini-PROTEAN 3 Gels (6)
Appendix B Guidelines for Gel Casting (7)
Section 1
General Information
1.1 Introduction
The Mini-PROTEAN 3 multi-casting chamber is ud to cast as many as twelve
0.5–1.5 mm thick gels simultaneously. The gels can be ud in both the Mini-PROTEAN 3
cell and the Mini-PROTEAN 3 Dodeca™Cell. After preparation, the gels can be stored up to
2 weeks at 4 ºC for future u.
1.2 Specifications
Materials of construction
Clamps Glass filled polycarbonate
Casting chamber, aling plate Molded polycarbonate
tubing
Gasket Silicone
Overall size 10 cm x 10 cm x 16 cm
g
Weight 500
Compatible glass plates Mini-PROTEAN 3 glass plates
Note: Mini-PROTEAN 3 multi-casting chamber components are not compatible with chlorinated hydrocarbons (e.g. chloroform), aromatic hydrocarbons (e.g. toluene, benzene) or acetone. U of such solvents voids all warranties. To insure best performance of the multi-casting chamber, become fully acquainted with the instructions before u. All components should be cleaned with a suitable laboratory detergent (Bio-Rad’s Cleaning Concentrate, catalog number 161-0772), rind thoroughly with distilled water, and dried before u.
Section 2
Loading the Chamber
1.Loon the clamp screws, remove the aling plate, and place the open casting chamber
body face up on the benchtop. The thumbscrews should face the ceiling.
2.Start by placing a paration sheet into the chamber so that it ats at the bottom. (Be
sure to remove the protective film from the paration sheet prior to u.)
3.Place a Spacer Plate (spacer side up) on top of the paration sheet.
4.Place a Short Plate on top of the Spacer Plate. Make certain that each addition is ated
at the bottom of the chamber.
5.Place a paration sheet on top of the Short Plate to complete one gel sandwich.
6.Repeat steps 3–5 until you have prepared the desired number of gel sandwiches.
德龙咖啡机使用7.Take up the remaining space in the chamber with acrylic blocks so that the sandwiches
will be held firmly in position when the aling plate is in place. When the chamber is
almost filled, Short Plates and, finally, the paration sheets can be ud to fill the
chamber flush to the top. If more than one glass plate is ud to take up the space, inrt
a paration sheet between the plates to simplify paration after polymerization.
Note: To insure a good al, the entire stack should be made as flush as possible to the top of the casting chamber, and not extend beyond it. If you overfill the chamber, monomer solution may leak out during pouring, and glass plates in the stack may break.
8.Seat the gasket firmly in the notch in the aling plate.
9.With the clamp screws looned, slide the aling plate under the clamps of the casting
chamber, being careful not to disturb the stack. The inlet port should match the groove at
the bottom of the chamber. Gradually tighten the screws in a random fashion until tight.
10.Stand the casting chamber up, and place it on a level surface. Do not tip the chamber
upside-down at this stage.
Section 3
Casting Gels
Single percentage gels can be prepared by introducing monomer from either the top or the bottom of
the chamber. Gradients must be introduced from the bottom.
The first time gels of a certain thickness are cast, it is necessary to empirically determine the required volume of acrylamide. Asmble the stack as outlined in Section 2, and inject a measured volume of water through the stopcock. Prepare this volume (+5 ml) of acrylamide.
Note: Wear rubber gloves while performing the following procedure to prevent accidental exposure to unpolymerized acrylamide, which is a neurotoxin.
3.1 Casting from the Top (Non-Gradient Gels Only)
1.Attach the stopcock valve to the inlet port of the casting chamber. Make sure that the
valve is in the clod position.
2.Mark a level on the chamber (with tape or a pen) at the desired paration gel length,
measuring from the bottom.
3.Combine all reagents except the initiators (usually APS and TEMED), and degas the
solution under vacuum for at least 15 minutes.
4.After degassing, add initiators to the gel monomer solution, and introduce the monomer
into the gel sandwich clost to the aling plate. A simple way to do this is to flow the
solution down the middle of the Spacer Plate of that sandwich using a pipet or a large
syringe. The groove on the bottom of the casting chamber will equilibrate the solution to
each of the gel sandwiches. Monitor the filling process by obrving the level of
solution rising on the sandwich furthest from the aling plate. Stop when you reach the
desired gel height.
5.Overlay the gels (if a stacker is required), or inrt the comb immediately.
6.Allow enough time for complete polymerization of the parating gel before removing the
中国历史朝代歌overlay solution (about 1 hour).
圆锥曲线公式
7.Prepare the stacking gel monomer solution as before and apply to the gels one at a time.
8.Inrt a comb into each gel sandwich. To minimize bubble formation, inrt the comb at
an angle.
Note: You may also cast single percentage gels from the bottom. U the gradient former, but just add the single percentage solution into both rervoir chambers.
3.2 Casting Gradient Gels from the Bottom
The inlet port on the Mini-PROTEAN 3 multi-casting chamber is ud for casting linear and convex acrylamide gradient gels. Refer to the gradient maker instructions for preparation of gradient solutions and proper operating techniques. The Model 485 Gradient Former (catalog number 165-4120) is recommended for u with the Mini-PROTEAN 3 multi-casting chamber.
An inexperienced ur should practice all steps ahead of time so that the procedure is completed quickly.
1. Place the gradient former on a magnetic stir plate and add a magnetic stir bar to the mix-
ing chamber labeled "light". Attach the luer fitting to the stopcock valve on the inlet port.
Run a piece of Tygon®tubing (1/8" ID Tygon tubing works well) from the gradient for-
mer to the luer fitting on the multi-casting chamber.
2. Determine the volume of monomer needed (e Appendix A).
3. Combine all reagents except the initiators, and degas the solution for 15 minutes.
4. Immediately prior to pouring, add TEMED and APS to both solutions, mix gently, and
pour the appropriate monomer solutions into the gradient chambers. (Consult the Model
485 Gradient Former instruction manual for complete instructions.) The light solution
(the one with the lower acrylamide concentration) should be placed in the mixing cham-
ber labeled "light", and the heavy solution in the rervoir chamber labeled "heavy".
5. Turn on the stirring bar in the mixing chamber, open the tubing clamp of the gradient
maker and the stopcock valve of the casting chamber, and pour the gels.
Note: If gravity flow isn't fast enough, u a peristaltic pump to pump the entire t of gradients within 10 minutes. If it is not possible to complete the operation in 10 minutes from the time initiators are added, then it might be necessary to reduce the amount of initiators (u ½ the amount of TEMED) to slow polymerization. The gradient should be poured as quickly as possible, without mixing the gradient solution in the casting chamber. Section 4黄疸是什么引起的
Chamber Disasmbly and Storage of Gels
Note: Wear rubber gloves while performing the following procedure to prevent accidental exposure to unpolymerized acrylamide, which is a neurotoxin.
1.Allow gel solution to polymerize for at least 1 hour.
2.Unscrew the clamp screws and remove the aling plate carefully.
3.Remove the gels one at a time from the stack. Separate the gel sandwiches from the acrylic
blocks and plates ud as space-fillers.
4.Rin off the tops of all the gels thoroughly with distilled water. Trim off excess
acrylamide around the glass with a razor blade. Wash off any pieces of excess acrylamide
prent with distilled water.
5.Store the gels upright in a tightly aled container or zip-lock bag. Add a few milliliters
of 1X gel buffer (identical to the buffer in the gel) to the bottom of the container and to
the tops of the gels to prevent them from drying out. Store tightly aled at 4 °C. Note: If
a stacking gel is required, consult Appendix B.
典韦字6.Clean the entire casting chamber thoroughly with distilled water. Residual acrylamide in
the stopcock valve and inlet port can be removed using a paper clip or a syringe needle.
