Question

1. Blend cells to DNA?) Keep in and release the - (Which organelle holds the solution to maintain water balance (osmotic balance). the mixture to pellet nuclei. the buffer solution, including the rest of the cell contents other than nuclei. 4. Resuspend the pellet of nuclei with 5. Check your progress: add a drop of stain to your suspension to stain and see the tiny nuclei under your microscopes. 6. Add EDTA to _the nuclear membranes. 7. Add SDS, a detergent, to the nuclear membranes. 8. Check your progress: add a drop of stain to your suspension to stain and see the "fluffy" DNA under your microscopes. This loose, readable form of DNA is called 9. Add NaCl to remove any , by interfering with protein- nucleic acid interactions. 10. Slowly add DNA is less soluble in than in water, so DNA is easier to remove from - (All 3 blanks are the same in this step.) 11. Carefully with a stirring rod, just below the the two layers and the DNA will out so that you can wind it up on the rod. 12. Once you have purified your DNA, you will run an experiment to check that what you have is truly DNA. You will set up three test tubes to do so. What is the negative control for this experiment? 13. What is the positive control for this experiment? 14. What is the chemical which turns blue when mixed with DNA and heated? of

Answer 1

1. disrupt cell membrane, nucleus, Tris buffer

2. Centrifuge

3. Decant or Discard

4. micropipette

5. Haematoxylin

6. destabilise and lyse

7. solubilise(the proteins and lipids of)

8.DAPI (fluoroscent stain)

9. DNA associated proteins (such as histones)

10. Isopropanol (in all three blanks)

11. Collect the DNA, interface, spool

12. To check whether your sample has DNA or not, take three test tubes and put following contents in these test tubes:

Test tube 1- Water+all PCR components (shows negative control)

Test tube 2- DNA +all PCR components(Sample)

Test tube 3- a DNA template + all PCR components. (positive control)

After agitating for specified time, run each one on the agarose gel and use bromophenol blue dye.

Negative control shows no amplification during PCR due to absence of DNA. If sample show band same as the Test tube 1 that means there is no DNA.

13. Positive control means the DNA is present in sample and getting amplified during PCR. If sample shows band same as Test tube 2, DNA is present.

14. Bromophenol blue.