- Attach Your Table Showing The Migration Distances For The Mw Markers 1 Kb Ladder Pcr Fragment And Vector Both Diges 1 (410.05 KiB) Viewed 41 times
- Attach Your Table Showing The Migration Distances For The Mw Markers 1 Kb Ladder Pcr Fragment And Vector Both Diges 2 (164.65 KiB) Viewed 41 times
- Attach Your Table Showing The Migration Distances For The Mw Markers 1 Kb Ladder Pcr Fragment And Vector Both Diges 3 (384.53 KiB) Viewed 41 times
- Attach Your Table Showing The Migration Distances For The Mw Markers 1 Kb Ladder Pcr Fragment And Vector Both Diges 4 (280.2 KiB) Viewed 41 times
- Attach Your Table Showing The Migration Distances For The Mw Markers 1 Kb Ladder Pcr Fragment And Vector Both Diges 5 (375.66 KiB) Viewed 41 times
Attach your TABLE, showing the migration distances for the MW markers 1 kb ladder), PCR fragment, and vector, both digested and undigested. You should have calculated the sizes for each band you detected on your gel (using the standard curve you generated from the MWM). Make sure these sizes are entered on the TABLE. Attach your log-linear graph with standard curve.
Use Loglinear graph paper to calculate the size of each band in your restriction digests (lanes 3-6). 1. Look at the picture of the "1kb ladder" (Fig. I-18), and, using that as reference, assign a size for each of the bands in lane 2 (the sizes of the top and bottom bands are provided in the Table). Put these values into the Table provided. 2. Using the ruler photographed with your gel, measure the distance traveled for each fragment on the gel (all the bands in the 1kb ladder AND all the bands in each of the other lanes of the gel) 3. Enter the "distance traveled" from the "well" where you deposited each sample for each band in each of the gel lanes (lanes #2,#3,#4,#5, #6) in the Table provided. As an example, uncut plasmid in lane #3 MAY have 3 different bands. You'll give their distances traveled with the top band being the one dosest to the gel origin. Your distances should be accurate to 0.1 cm(1 mm). 4. First generate a STANDARD CURVE from the information you have for your " 1 kb ladder." Note that for each band in this lane, you know BOTH the size of the fragment AND the distance that the fragment traveled. Assign a "dot" on the log/linear graph for each fragment in the "1kb ladder", indicating distance traveled on the X-axis (horizontal axis = linear scale) and the size of the fragment on the Y-axis (vertical axis = logarithmic scale). You should plot 20 dots for the "1kb ladder". When all dots are placed, draw a straight line from each dot to the next. You have now created your STANDARD CURVE (see an example in Fig. I-19). 5. Now determine the size of all the other fragments on your gel. You have already determined the distance traveled for each fragment. Use a pencil to find the place on the X-axis corresponding to that distance. Draw a straight line up (vertical line) from that place on the X-axis unti it intersects with the STANDARD CURVE. At that point, draw a straight, HORIZONTAL LINE to the left until you reach the Y-axis. The point of intersection on the Y-axis = size of the fragment.