please how can the concentration of the DNA in the mixture b

please how can the concentration of the DNA in the mixture be calculated

Laboratory week 2 Preparation of lambda phage genomic library objectives: 1. In this laboratory session you will prepare a genomic library of DNA In order to accomplish this goal you will cleave DNA as well as pUC18 DNA with the restriction enzyme, EcoR 1. The DNA fragments generated via this digestion will then be ligated into the pUC18 linearized plasmids to create the lib recombinant plasmids containing different segments of the phage genome Laboratory Schedule 1. Digest DNA and pUC18 with EcoRI 2. Prepare an agarose gel for use in next week\'s laboratory session. 3. Ligate your EcoRi generated DNA fragments to EcoRI digested pUC18 Procedures Restriction Digestions l obtain two microfuge tubes labeled and \"pUC18\" 2. A be in the tube marked \"a\" ul (00ug/ml) of pUC will be in the tube marked \"pUC and 20 18 3. Add 40 ul of EcoRI- Buffer (20 enzyme units where one unit is defined as the amount of enzyme required to digest lug DNA in one hour at 370C in the appropriate buffer) to each tube and mix. The buffer is composed of 50mM Tris-HCl (pH 8.0), 10mMMgCl2, and 100mM Naci. 4. Incubate the tubes at 37 C for 50-60 minutes in a heating block. 5. At the end of the incubation period, label two tubes marked \"h-E\" and \"pUC18-E (E-electrophoresis) and place 3 ul of electrophoresis sample buffer into each tube from 10ul of the digestion from tube into tube \"pUC18-E\" and 10ul of the digestion tube into tube \")-E\" 7. Store theses tubes in the freezer for gel electrophoretic analysis during next week\'s laboratory This analysis should reveal whether EcoRi actually cleaved your DNA sample 1. Label a microfuge tube \"L\"(for ligation) and add 5 ul of the EcoR1-cut pUC18 DNA. (contained in tube \"pUC18\") and 20 ul of EcoRI-cut DNA (contained in tube 2. the tube in a 70 Cheating block \"n\' o the tube Heat 3. Place the for 10 minutes in order to inactivate the EcoRI enzyme tube on ice for 5 minute to allow its contents to cool and then add 50 uDof cold DNA Ligase-Buffer solution. This solution will contain 1.5 units of T DNA Ligase in a 2x ligase buffer (1x buffer is composed of 50 mM Tris-HCI (pH 7.6), 10 mM MgC12, 1 mM ATP, 1 mM dithiothreitol, 5% (w/v) polyethylene glycol 8000). Ligase catalyzes the formation of a phosphodiesterbond between adjacent 3\' hydroxyl and 51 phosphate termini in double stranded

Solution

iN THIS EXPERIMET,

the stock solution of DNA ihas been prepared and from that stock solution, required amount of DNA has been taken.

stock which is prepared is 300 microgram per ml. it means that In 1 ml of buffer or water, 300 Microgram of DNA is mixed and from this stock solution, 20 microliter of DNA has been used in the digestion experiment. 300 microgram will be equal to 0.3 mg.