Carefully remove the comb and tape surrounding the plate and transfer the gel plate to the electrophoretic tank such that wells are towards the cathode 4. Pour 1X TAE buffer into the tank until the gel is completely submerged. Connect the electrode to the power supply 5. Load the plasmid DNA preparation and standard DNA marker into separate well with help of a micropipette or a syringe 6. Turn ‘ON’ the power supply and run at 100 V (10-15 mA).
Abstract The transformation principle suggests that bacteria use DNA as their genetic material and are able to exchange their genetic material via a process of transformation. Griffith had theorised the concept of the transformation principle using two strains of bacteria and studied their ability to recombine. Avery and MacLeod followed his studies and suggested DNA was sensitive to DNase, and that the enzyme would destroy the bacteria's ability to exchange genetic material and transform into a new strain. This was then tested in the labs at Wits by second year students where they studied the transformation of ampicillin sensitive E. coli to ampicillin resistant E. coli. The results obtained there were similar to those of Avery and MacLeod,
The formation of a dye-iodine complex will occur in the cytoplasm. Then, it was flooded with ethanol and washed immediately. It is where the process of decolorization occurs. It should not be prolonged as it can over decolorized and immediate washing would sometimes cause under decolorized smear and. Finally safranins were flooded on slide for 30 seconds and rinse it with tap water and the slide must be dried using a blotting paper before viewing and examine in the
Approximately 2 gm, nearest to 0.1 mg, oven dried cornhusk fibres, were weighed out accurately in weighing bottle and transferred to a 100 ml beaker. 40 ml of cold (10-15˚C) 72% sulphuric acid was added gradually to the fibres in small increments while stirring the mixture and macerating the fibres with a small glass rod. The beaker was kept in a bath at 2 ± 1˚C for dispersion of material. After the specimen was dispersed, beaker was covered with a watch glass and kept in a bath at 20 ± 1˚C for 2 hours. Mixture was stirred frequently to ensure complete
Waist circumference was measured at the mid-point between the lower borders of the rib cage and the iliac crest. Semen Analysis: Standard semen analysis was performed according to the guidelines of the fifth edition of the WHO laboratory manual for examination of human semen.19 Participants were asked to deliver a semen sample (by masturbation) into a sterile plastic container after 2-7 days of sexual abstinence. The samples were left to liquefy in a 37°C incubator, and were analyzed within one hour of delivery. Standard procedures included
The column was centrifuged empty at full speed for 1 min to remove any residual ethanol. The RNA was eluted by adding nuclease-free water into the column and centrifugation of 1 min at full speed. The yielded RNA was checked by nanodrop (Thermo) and the integrity of the RNA was assessed in the Experion automated electrophoresis station
¾). Placed the cuvette sample in the Sprectrovis. After each run, the temperature of each sample was collected (to nearest 0.1°C). Disposed of the sample solution, cleaned the cuvette with DIW and repeated the latter procedure using the correct volumes for each new run from Table 1.
1.Polymerase chain reaction (PCR): Polymerase chain reaction is a method of DNA or RNA amplification . The PCR method allows millions of copies to be created from a very small DNA section. The PCR methodology was developed in 1983 by Kary Mull , who in 1993 received a Nobel Prize for Chemistry with Michael Smith PRINCIPLE & PROCEDURES: 1.DNA denaturation. Once the DNA has been isolated and purified from the cell, a PCR assay can begin. Uncleaned DNA can also be used for PCR, but it is ineffective.
22.5 g of plate count agar powder was dissolved in a litre of sterile distilled water on the hot plate 2. pH of the solution was adjusted to 7.0 ± 0.2 by adding NaOH or HCl and was immediately transferred into the Schott bottle to be autoclaved at 121 ° C for 15 minutes 3. Prepared medium was stored in 4° C chiller Lauryl Sulphate Broth
The first well was pre-loaded with DNA ladder and labeled as 1 microliter kb DNA ladder. Next one microliter of DNA was mixed with one microliter of loading dye using a pipettor and loaded into the well. The same mixing process was completed for the PCR product, using one microliter of PRC product and one microliter of loading dye. For the purposes of this experiment, the DNA product was loaded into well six and the PCR product was loaded into well seven. Initially DNA was loaded into well five, however gel was pierced so samples were moved one well to the right.