Katpak:m1d2

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
Line 1: Line 1:
1. Take the log10 of the length of each molecular weight marker you can identify on your agarose gel photograph. Graph the log10 of their length on the y-axis versus the distance they migrated from the well on the x-axis, measured in mm using a ruler and the picture of your agarose gel. An example of such a graph is found in the introduction to today’s experiment. Use the equation of the line from your graph to determine the size of your M13K07 backbone (use the band in the lane in which you loaded the cut DNA). How does this measurement compare with the predicted size?
1. Take the log10 of the length of each molecular weight marker you can identify on your agarose gel photograph. Graph the log10 of their length on the y-axis versus the distance they migrated from the well on the x-axis, measured in mm using a ruler and the picture of your agarose gel. An example of such a graph is found in the introduction to today’s experiment. Use the equation of the line from your graph to determine the size of your M13K07 backbone (use the band in the lane in which you loaded the cut DNA). How does this measurement compare with the predicted size?
 +
{| border="1"
{| border="1"
Line 35: Line 36:
| 2.70 || 52
| 2.70 || 52
-
|}
+
|}  
 +
 
 +
[[Image:M2d2gel.JPG|400px]]
 +
 
 +
The cut fragment was at 19mm. Thus, from the equation of the line on the graph above, y=3.9421 and the size of the fragment is ~8700bB, which is comparatively close to the expected size of M13K07 of 8669bB. 
 +
 

Revision as of 11:39, 19 September 2007

1. Take the log10 of the length of each molecular weight marker you can identify on your agarose gel photograph. Graph the log10 of their length on the y-axis versus the distance they migrated from the well on the x-axis, measured in mm using a ruler and the picture of your agarose gel. An example of such a graph is found in the introduction to today’s experiment. Use the equation of the line from your graph to determine the size of your M13K07 backbone (use the band in the lane in which you loaded the cut DNA). How does this measurement compare with the predicted size?


Data
Marker Log(bp) Distance (mm)
10 4 19
8 3.903 21
6 3.78 23.5
5 3.70 25.5
4 3.60 27.5
3 3.48 31
2 3.30 35
1.5 3.17 39
1 3 49
0.5 2.70 52

The cut fragment was at 19mm. Thus, from the equation of the line on the graph above, y=3.9421 and the size of the fragment is ~8700bB, which is comparatively close to the expected size of M13K07 of 8669bB.


2. How many plaques do you expect if you plated 10 ul of a 10^-8 dilution of phage, if the titer of phage was 10^12th plaque forming units/ml? How many plaques would you expect if you tested the phage stock on strain DH5?

2. (10^12PFU/mL)(10^-8)*(10uL)/(10^3uL/1mL) = 100. Thus, you would expect to see 100 plaques.

3. E-mailed.

4. The oligonucleotide you are adding to p3 uses traditional genetic engineering ("recombinant") techniques. These are powerful and precise ways to move single genes from one organism to another and to make useful chimeric protein products like the one you are now working on. Synthetic biology is a newer approach to programming cells. Please read one (or more!) of the following articles and then write a paragraph exploring the legitimacy of the following statement: "synthetic biology is about engineering while genetic engineering is about biology."

Personal tools