Dna digestion and electrophoresis

In general terms, smearing is when you have many bands together close enough in size that you cannot distinguish between adjacent bands i.

restriction digestion principle

If this experiment was performed without significant error, the likely explanation is that a 4-base cutter was used. This is called a ladder.

Restriction enzyme digestion lab report

Remember that uncut DNA can be found in 3 forms, and the slowest of these is linear. Does the data seem reasonable? All fragments in this lane are generated by digestion of one particular DNA, so fragments are present in equimolar amounts and the brightness of the bands corresponds to their well-known lengths. DNA samples for loading into the wells "slots" of the gel are prepared by addition of a tracking dye e. Slide gel easily into staining tray labeled with your group name. It is unlikely that one could see 25 individual fragments of such a small size, and the smearing pattern is probably what would be detected. It is clear that in lane 2 two fragments are present in non-equimolar amounts the upper band must contain longer DNA molecules, but is less intense than the lower band.. Carefully remove the casting tray and keep it horizontal until you are ready to put into the plastic container. This step eliminates any air in tip. If it is present, put the sample back and begin again.

The gel is now ready to load with DNA. You will need to remember the order of your tubes since there is NO way that the gel can be marked.

Dna digestion and electrophoresis

Smaller DNA molecules migrate faster than large ones. This is because there is more nanograms of DNA in 3 than in 2 the number of molecules in 3 must be much higher than in 2.

Lambda dna restriction digest and electrophoresis lab

Do NOT move tray while agarose is solidifying. Not in this class. Is there anything significant about 6. Cooling down to room temperature results in the slow formation of a solid gel when the concentration of agarose is between 0. Yes, it's about half of our original sample. Pool and mix reagents by tapping the tube bottom on counter a couple of times or use a microcentrifuge. Always touch pipette tip to side of tube to dislodge any small amount stuck to tip. Size markers may be co-electrophoresed with DNA samples, when appropriate for fragment size determination.

Looking at the gel you see one band approximately 6. Casting agarose gel Set gel-casting tray into the tray apparatus, screw tight, and insert well-forming comb into space marked with red line.

agarose gel electrophoresis

It is clear that in lane 2 two fragments are present in non-equimolar amounts the upper band must contain longer DNA molecules, but is less intense than the lower band.

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Restriction Endonuclease Digestion and Agarose Gel Electrophoresis of DNA