150 volts until orange dye have migrated an appropriate distance about 5 6 cms 10 excess agarose can be stored at room temperature and remelted in a microwave and can be used again a all reaction give pcr product of 800 base pair for internal control this proves pcr reaction is working fine and there is no inhibitor of pcr in the clinical sample b only chlamydia trachomatis positive sample give an extra band of 368 bp as is evident from the dna size marker lane 1 take 500 u03bcl of collected sample cocktail in a fresh microcentrifuge tube 1 5 ml and centrifuge at 15 000 g for 30 minute at 4 u00b0 c 2 discard the supernatant carefully without disturbing the pellet 3 suspend the pellet from step 2 in 50 u03bcl of te solution 4 now incubate the sample at 100 u00b0 c for 10 minute 5 spin at 10 000 rpm for 2 minute 6 collect the supernatant carefully in separate tube designated as sample dna all other steps will be as mentioned in the detailed assay sample is collected as a swab and is dipped in the vial containing sterile 1m1 transport medium and stored at 4 u00b0 c for several hours or in freezer for a maximum time up to one week 1 take 500 u03bcl of collected sample cocktail in a fresh microcentrifuge tube 1 5.