please help me fill the table. Thank You.(ASAP)
NaOH=M1V1=M2V2
=(2M)(1ML)=(0.2)(10ML)
SDS=(10)(1)=(1)(10)
=M1V1=M2V2
MATERIALS Sample 100mL Escherichia coli (E.coli) culture Apparatus Latex gloves 1.5mL microcentrifuge tubes Centrifuge, refrigerated, with rotor for 1.5mL microcentrifuge tubes P1000 micropipette and sterile tips Incubator shaker 20C freezer Universal bottle pH meter Glassware and labware Flake ice Ice box Chemicals Sterile deionized water (sdl2O) Sterile saline solution (0.9%w/vNaCl in sdH2O) Glucose Tris-Cl Ethylenediaminetetraacetic acid (EDTA) Sodium hydroxide (NaOH) Sodium dodecyl sulphate (SDS) Potassium acetate Glacial acetic acid Ethanol 96%v/V,70%v/v (cold) Nutrient broth media Tris/Borate/EDTA (TBE) buffer 1 RNase (optional) Solution A2 (Glucose 50mM; Tris-Cl 25mM; EDTA 10mM ) Solution B3(NaOH0.2N ' SDS 1% w/v) Solution C4 (Potassium acetate 5M, glacial acetic acid) ROCEDURE 1. The bacteria culture was shaken gently in the shake flasks and 1mL was transferred to a sterile 1.5ml microcentrifuge tube. The tube was labeled as A. 2. The tube was centrifuged at 10,000rpm for 1 minute at room temperature. 3. The supernatant (media broth) was poured off immediately and the excess supernatant fluid was drained by inverting the tube over a clean tissue paper. The undisturbed pellet (bacterial cells) was left behind at the bottom of the tube. 4. Steps 1-3 were repeated twice in the same tube. The tube was filled again with more bacterial culture. 5. The pellet was washed by adding 1mL of sterile saline solution and mixed by the bottom of the tube was gently flicked. 6. The tube was centrifuged at 10,000rpm for 1 minute, then the supernatant was poured off and left behind the pellet. 7. The pellet was resuspended with 200L of solution A. The bottom of the tube was gently flicked and left for 5 minutes at room temperature. 8. The mixture was added with 400L of solution B, mixed by the tube, was gently inverted five times and put in an ice bath for 5 minutes. 9. After 5 minutes, 300L of solution C, mixed by the tube, was gently inverted five times and put in an ice bath for 5 minutes. 10. The tube was centrifuged at 12,000rpm for 11 minutes. 11. A P1000 micropipette was used to carefully transfer the supernatant of the mixture in tube A into a new 1.5mL microcentrifuge tube that was sterilized, without disturbing the pellet. This tube was labeled as A1. 12. The tube A was discarded into a plastic disposal bag. The tube A1 was used for further analysis. 14. The tube A1 was kept in the 20C freezer for about 10 minutes. 15. The tube A1 was centrifuged at 12,000rpm for 10 minutes. 16. The supernatant (ethanol) was poured off immediately and the excess supernatant fluid was drained by inverting the tube over a clean tissue paper. The undisturbed pellet (plasmid DNA) was left behind at the bottom of the tube. 17. A P1000 micropipette was used to add 500L of 70% cold ethanol into the tube and miced by the bottom of the tube was gently flicked. 18. The tube A1 was centrifuged at 12,000rpm for 5 minutes. 19. The supernatant (ethanol) was poured off immediately and the excess supernatant fluid was drained by inverting the tube over a clean tissue paper. The undisturbed pellet (plasmid DNA) was left behind at the bottom of the tube. 20. The tube cap was opened and placed in the laminar air flow for drying. 21. After the tube is completely dried, 20L of sdH2O and I L of RNase was added and mixed by the bottom of the tube was gently flicked. 22. The tube at 20C was kept for about one week to preserve the extracted plasmid DNA for electrophoresis. Table 5.1 Concentration values
