23 Jul WHY DO THE DNA BANDS APPEAR BRIGHT AGAINST A DARK BACKGROUND?
A restriction digest is prepared using the two restriction enzymes Kpn I and SacI on a fragment of Show more A restriction digest is prepared using the two restriction enzymes Kpn I and SacI on a fragment of DNA (like a PCR product for example). Undigested the fragment was 9000bp (or 9kb) in length (See table 1). The digests would result in the following sized fragments assuming complete digest and no star activity DNA Size of Fragments uncut DNA 9000 DNA cut with Kpn I 7000 and 2000 DNA cut with Sac I 4500 and 4500 DNA cut with Kpn I and Sac I 4500 2000 2500 After you complete the digestion reactions you add some loading dye then run your digest samples on a gel which has been stained with ethidium bromide. You observe the gel under UV light in order to confirm if your digest worked and if it successfully produced the predicted sized fragments (Figure1 below). Use table/figure 1 to answer the following: 15 points Figure 1: 1. Why was a sample of size markers loaded (in the far left well of the gel)? 2.
A restriction digest is prepared using the two restriction enzymes Kpn I and SacI on a fragment of Show more A restriction digest is prepared using the two restriction enzymes Kpn I and SacI on a fragment of DNA (like a PCR product for example). Undigested the fragment was 9000bp (or 9kb) in length (See table 1). The digests would result in the following sized fragments assuming complete digest and no star activity DNA Size of Fragments uncut DNA 9000 DNA cut with Kpn I 7000 and 2000 DNA cut with Sac I 4500 and 4500 DNA cut with Kpn I and Sac I 4500 2000 2500 After you complete the digestion reactions you add some loading dye then run your digest samples on a gel which has been stained with ethidium bromide. You observe the gel under UV light in order to confirm if your digest worked and if it successfully produced the predicted sized fragments (Figure1 below). Use table/figure 1 to answer the following: 15 points Figure 1: 1. Why was a sample of size markers loaded (in the far left well of the gel)? 2. Why do the DNA bands appear bright against a dark background? 3. If in fact the digest was successful explain why is there only one band appearing for the Sac I digestion when you predicted a product of two 4500bp fragments? Show less
3. If in fact the digest was successful explain why is there only one band appearing for the Sac I digestion when you predicted a product of two 4500bp fragments? Show less
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