• Learn operation of Electrophoresis tank and Electrophoreis Power supply.
• Read following documents

Electrophoresis is a comprehensive term that refers to the migration of charged solutes or particles of any size in a liquid medium under influence of an electrical field.

• Document and revise common laboratory informations related to the examinations

1. 1% Agarose gel
2. Hippurate buffer pH 8.8
3. Amido black stain
4. CBB
5. Methanol
6. 5% glacial acetic acid
7. BPB(bromophenol blue) dye

1. Before applying the gel on the slide wash 2 slides(Non siliconized slide on which gel will be applied and siliconized slideSiliconization of Glassware will be used as a slider) and dry it with tissue roll. siliconized side is the one with red mark.
2. Place 2 slides on a levelled surface as shown in above figure.Cut a rectangle of slide size from a x-ray plate, cut a four boardered frame from a x-ray plate, stick it to the border of non siliconized slide through feviquick. so the thickness of gel is same as thickness of x-ray plate. (take only a drop of feviquick on four corner.
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5. Prepare 1% Agarose gel.
6. Then pour hot agarose gel on non siliconized slide carefully.Make a wedge of siliconized slide with red mark downward side on non siliconized slide containing agarose gel(as shown in figure).
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8. Carefully press the siliconized slide over the non siliconized slide,so that air do not get trapped in between two slides. Do not give so much pressure otherwise air will come in between 2 slide on removing pressure
9. Gel will be ready to use after 10-15minutes.
10. 

1. Bring used histopathology blade from histopathology department after permission of in-charge of that respective department.
2. By making 2mm slot at every 5mm distance to prepare comb.
3. Wrap 3 layers of cellotape at each end of applicator comb so that level of applicator comb will remain feww milimeter higher from the gel so can prevent through and through cutting of gel and spreading of samples beneath the gel.
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5. If you are using slide with sticked X-ray film border, then use applicator which has slightly short ends.( Such applicator is made by Dr. SMP, Please confirm before use)

1. Allow the samples to clot completely before centrifugation.Take 200 microliter of serum in each eppendroff cups.
2. For control—>take 200 microliter of plasma & 200 microlter of normal serum in each eppendroff cups. Add 20 microliter of BPB(bromophenol blue) dye

in each eppendroff cups mix it thoroughly.

1. Hold the prepared gel slide in your one hand.
2. With thumb of one hand,fix the x-ray film of non siliconized slide,slowly and carefully remove the siliconized slide by sliding it with other hand.
3. Take care that gel do not get broken down or wrinkled while removing the siliconized slide.
4. Loading sample over applicator:
   1. 
   2. Taking sample directly over applicator will result in loading of excess sample over gel.
   3. So first keep magnet attached applicator comb on a clean microscope glass slide to remove excessive sample application over gel.
   4. Take 2ul of sample with 2ul pipette from prepared sample cup and apply it over applicator.
   5. After slight elevation of applicator,Remove the microscope glass slide. If sample is applied on both side of applicator then remove it through tissue paper taking care not to remove sample from both side.
5. Loading sample over Gel:
   1. put prepared gel slide beneath the applicator.
   2. The applicator is put over one end of gel loaded slide about 1-1.5 cm away from margin.
   3. Wait for 2-3 minutes,allow the samples to get completely absorbed within gel otherwise samples will spread.
   4. Then remove the applicator carefully.

1. Clean the electrophoresis tank with DI water and dry it with tissue paper before use.
2. Put the electrophoresis tank on a levelled surface. Fill the hippurate buffer in the two compartments.
3. After successful sample application,put the gel slide in electrophoresis chamber.
4. Connect the gel with buffer through strip of filter paper(wick). & cover the chamber with its lid.
5. Connect the electrode on the side of slide containing sample well to the cathode (-) & other electrode to the anode of the power supply.
6. Set voltage at 500 and Set current at maximam & run for 10 minutes.After 10 minutes,switch off the power supply & remove the slide.

1. First remove water around the proteins and fix the protein by putting the slide in methanol solution for 10 minutes.
2. fix the gel by heating in microwave for 1.5 minutes. Allow the slide to be cool.Then place the slide in a staining solution CBB or Amido black stain for 2 minutes.
3. Remove the excess stain by putting in the destaining solution 5% glacial acetic acid for 15 minutes.Allow the slides to dry at room temperature.Analysed the results.

1. label the slide with appropriate sample id according to sample application.
2. Keep the labeled slide on RICHO-2000L2 for scanning as shown in below figure.
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4. Set the “Resolution” around “600dpi” as shown in figure.
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6. Set thr scanner as shown below
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8. Press on “start”.
9. Press on “#” for sending the file which is scanned.
10. Open the “file manager” in resident room computer.
11. Open the “File system”—–>Click on “Scan” folder—–>Do right click on the image which is scanned—–>Open wih “GNU Image Manipulation Programme”.
12. Slide will be seen as shown in below figure after clicking on GNU Image Manipulation Programme.
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15. Do right click on that selected area—–>click on “Edit”—–>Click on “Copy”—–>Do right click on that selected area—–>click on “Edit”—–>Click on “Paste as”—–>Click on “New image”.New image is seen as shown in below figure.
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18. After Clicking on “export” below figure appear.click on “Export” also.
19. 

1. Go to Application menu—–>Click on web browser—–>Open 10.207.3.240
2. Do Login from your login ID.
3. Click on “Edit”—–>Click on “Manage attachment” as shown in below figure.
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5. 2 attachments should be done - 1st is “electrophorogram” and 2nd is “Interpretation” file.
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7. Do 2nd attatchment as shown in below figure.
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9. After click on “Browse”, attach the “interpretation file(txt file)”.

1. Usually the following 5 bands are seen starting farthest from the origin :
2. albumin—>alpha-1 globulin—>alpha-2 globulin—>beta globulin—>gamma globulin.