Lab Flow Chart #4 PDF

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Chan Park General Biology 1 Lab Dr. Pearlman Flow Chart Week #7 10/09/19 Electrophresis 1. Put on a lab coat and gloves. 2. Fit a clean comb into gel tray, and put the tray into the electrophoresis box and put the gel casting gates on the sides of gel tray 3. Add molten agarose slowly to the gel tray until the small tray is half-full and wait 15 min for the gel to harden 4. Label a clean 1.5mL tube for each sample with both the sample number and the words “for gel”. 5. Add 2uL of 6x gel loading dye to each "for gel" tube and then add 3 uL distilled water and 5 uL of the PCR sample to each "for gel" tube. Save the rest for DNA sequencing by placing it in the ice. 6. Obtain 10 uL DNA ladder with dye already added to it. 7. Remove the gates from the sides of the gel casting tray. 8. Fill the box with 1x TBE until it just covers the gel. 9. Carefully remove the comb from the gel. If gloves used to stabilize the gel, change it immediately afterwards 10. Wipe the barriers and comb with a wet tissue. And dry them before putting them away. 11. Draw a picture of which item you plan to load into which well of the gel and keep it in the safe place 12. Use a micropipettor to load each sample into a separate well of the gel. And then, put the cover on the gel box, connect the electrodes to the box and to the power supply. 13. Run the gel at 100 - 115 volts until the faster dye moves almost completely off the gel for about 30 minutes. 14. Check gel after 30 seconds to be sure that (1) there are bubbles forming near the electrodes, (2) the power supply is pulling current (switch the readout to amps or current to check), and (3) your sample is moving in the correct direction. 15. Obtain a large weight boat and label the rim with name. 16. Turn off the power supply and remove the top from the gel box. Then, lift the gel tray out of the box. Slide the gel out of the tray and onto the weigh boat. 17. Use the gel documentation system to visualize the gel. 18. Dispose of gel and gloves in the biohazardous waste container, hang the lab coat back in the designated area (unless something spilled), and compare your sample to the DNA ladder....