Talk:BE.109:DNA engineering/Examine candidate clones

From OpenWetWare

Jump to: navigation, search
  • In the future, I think it would be better to use 1 ng DNA instead of 5. The colonies were far too crowded to count on the 5 ng plate, which makes the homework assignment for tomorrow impossible.
  • Definitely better to use less DNA.
  • Could also plate less of transformation mix...
    • To complete the calculation, you can roughly estimate the crowded plate as 5000 colonies, though this could easily be 2X off in either direction. (Thanks to Yoon Sung for catching the error in the homework related to the plasmid's name...it should be pCX-EGFP) And if you'd like to see the class data again, here it is -- Natalie
    • For future reference, another option is to count the colonies in a sector of the plate (i.e. 1/8th or 1/16th ... draw lines on the plate like pie slices to divide into sectors) and then multiply by the number of sectors. This should lead to a reasonable approximation to the number of colonies. Of course, this only works if your colonies are actually distinguishable from one another. If you have a continuous lawn of cells, this won't work. Regardless, go with Natalie's suggestion for your homework assignment. -- Reshma (your Mod 3 TA chiming in).


Tuesday/Thursday

Green Purple Red Blue Pink Yellow
5ng lots lots 1005 1500 lots tons
no ligase control 1 7 5 0 0 0
bkb + ligase control 1 19 8 0 1 114
complete ligation mixture 21, 22 800, 850 75, 80 7, 13 65 89, 105

Wednesday/Friday

Green Purple Red Blue Pink Yellow
5ng lots lots lots lots lots lots!
no ligase control 400 0 1? 0 0 3
bkb + ligase control 0 8 0 1 0 3
complete ligation mixture 220, 250 22, 28 0, 0 0, 0 6, 5 44, 44
Personal tools