RE Digestion Field Trip Results Lodi High School Mrs. Bielec’s Group March 8, 2011
Lodi HS Student Data: Anna 1 2 3 4 5 6 Lane 1 Lane 2 Lane 3 Lane 4 Lane 5 Lane 6 10l Negative (No Enzyme) Control Sample ? E / H Lambda Digest Marker Team Anna: Observations: While not perfect, this is the gel that was used as the class model during the field trip. Lane 1 clearly shows a single band for the negative control sample. Lane 5 shows the banding pattern that we expect to see in the Digest Marker. Both of these controls look good! Lane 3 shows a robust banding pattern that matches the Lane 5 digest marker. This indicates that Lane 3 is the EcoRI/HindIII double digest. Lane 4 shows a HindIII banding pattern: The largest digest fragment is larger than the largest fragment in the Lane 5 digest marker. (~23,000bp vs. ~21,000bp) The smallest visible HindIII digest fragments (2,322bp and 2,027bp) are smaller than any fragment that one would find in an EcoRI digest (smallest EcoRI fragment is ~3,500bp) Lane 6 appears to be the PstI digest result: The largest digest fragment is smaller than the largest fragment in the Lane 5 digest marker (~21,000bp) Since PstI cuts Lambda phage DNA 28 times (relative to EcoRI’s 5 and HindIII’s 7), the Lane 6 digest more total visible fragments. Lane 2 has a small smudge, but no definitive bands. This could be due to a number of things, but is likely linked to some sort of pipette error in setting up the digest. If the previous observations are correct, then Lane 2 can be implied to be the EcoRI digest.
Lodi HS Student Data: Brooke 1 2 3 4 5 6 Lane 1 Lane 2 Lane 3 Lane 4 Lane 5 Lane 6 10l Negative (No Enzyme) Control Sample ? E / H Lambda Digest Marker Team Brooke: This team had four wells with an interpretable result. Observations: Lane 1 clearly shows a single band for the negative control sample. Lane 5 shows the banding pattern that we expect to see in the Digest Marker. Both of these controls look good! Lane 3 shows a HindIII banding pattern: The largest digest fragment is larger than the largest fragment in the Lane 5 digest marker. (~23,000bp vs. ~21,000bp) The smallest visible HindIII digest fragments (2,322bp and 2,027bp) are smaller than any fragment that one would find in an EcoRI digest (smallest EcoRI fragment is ~3,500bp) Lane 6 appears to be the PstI digest result: The largest digest fragment is smaller than the largest fragment in the Lane 5 digest marker (~21,000bp) Lane 2 appears to not have gotten any Lambda phage DNA. Lane 4 has what appears to be a smudge of DNA that bands at about 21,000 bp, but this isn’t enough to tell the difference between the EcoRI and EcoRI/HindIII double-enzyme digests.
Lodi HS Student Data: Harley 1 2 3 4 5 6 Lane 1 Lane 2 Lane 3 Lane 4 Lane 5 Lane 6 10l Negative (No Enzyme) Control Sample ? E / H Lambda Digest Marker Team Harley Observations: Lane 5 shows the expected pattern from the digest marker. No other lanes on this gel show DNA. It is likely that there was an issue with sample set-up in this group.
Lodi HS Student Data: Mitch 1 2 3 4 5 6 Lane 1 Lane 2 Lane 3 Lane 4 Lane 5 Lane 6 10l Negative (No Enzyme) Control Sample ? E / H Lambda Digest Marker Team Mitch Observations: Lane 5 shows, faintly, the banding pattern expected from the digest marker. Lane 1 shows, faintly, one large uncut band, which is what we expect from the negative control Lane 2 looks very similar to the negative control in Lane 1, indicating that it did not receive restriction enzyme. Lane 6 shows a very faint result. Since the largest fragment in Lane 6 is smaller than the largest fragment in the Lane 5 digest marker, it is reasonable to conclude that Lane 6 contains the PstI digest, even though the result is faint enough keep us from resolving the majority of the 29 fragments expected from a PstI digest of Lambda DNA. While DNA is clearly visible in both Lanes 3 & 4, it is difficult to interpret with certainty which of the remaining digests might be seen in them.
Lodi HS Student Data: Taylor 1 2 3 4 5 6 Lane 1 Lane 2 Lane 3 Lane 4 Lane 5 Lane 6 10l Negative (No Enzyme) Control Sample ? E / H Lambda Digest Marker Team Taylor: Observations: Lane 1 has one solitary band, which is what we expect to see in the negative control well Lane 5 has a set of DNA fragments. Though faint, they show a pattern that approximates the Lambda digest marker. Lane 6 appears to be the PstI digest result since the largest digest fragment is smaller than the largest fragment in the Lane 5 digest marker (~21,000bp) Lane 4 has one very bright band indicating that while DNA was placed into this digest tube, while restriction enzyme was not. Lanes 2 & 3 contain not enough DNA to see resulting fragments.