Question

Molecular Key Calculation - PLEASE NEED HELP TO CALCULATE IT!

BstAPI 179 Eco010912674 Aat ll 2617 Sspl 2501 Ndel 183 Ehel 235 Xml 2294 2486 Bcgl 2215 Scal 2177 146/ 236 MCS Tacz 77 469 bla (Ap) pUC18/19 Sap 683 2686 bp 852 Afl, BspLU11 806 1626 1466 Gsul 1784 Cfr101 1779 Eco311 1766 Eam11051 1694 rep (PMB1) 3. AWNI 1217 Using the map above what size band/s would you get on a electrophoresis gel after you have cut puc18 with Nde I?

1. You are setting up you own Master Mix for doing a Not1 restriction digest.

             In each digestion tube you will be adding 10 ul DNA and 40 ul of your Master mix.

             You are given 10 x Not1 buffer, 10 ug/ul acetylated BSA, Not1 restiction enzyme 10 units/ul and sterile water.

             How much of each of the 4 reagents would you use to make 500 ul of Master Mix.

             The 50 ul final digest mix should contain 1 x Not1 Buffer, 1 ug/ul acetylated BSA, and 2 units Not1 plus 10 ul DNA.

2. How would you make 1000 ml of a solution that is 20 mM MgCl₂, 10 mM CaCl₂ and 50 mM KCl from 0.5 M stocks of each of these chemicals?

Using the map above what size band/s would you get on a electrophoresis gel after you have cut puc18 with Nde I?

4. You dilute your plasmid DNA that you have purified 1 in 2 and then you measure OD₂₆₀ of the diluted DNA at 0.25 OD, what is the actual concentration of your purified DNA ( 1 OD 260 unit is equal to 50ug/ml DNA).
5. You are setting up your own PCR Master Mix. You are given 25 mM MgCl2, 10 X PCR Buffer, sterile water, 20mM dNTPS, 100mM primer mix and Taq polymerase at 7 Units/µl. How much of each of the 6 reagents and water do you need to add to make a 500 µl master mix with a final concentration of 2 mM MgCl2, 1 x PCR buffer, 2mM dNTPS, and 10 mM primer mix. If this mixture is for 10 samples, and each sample will contain 1 unit of enzyme, how much Taq do you need to add to your master mix?