Tubes 1-7, final Mg++ conc. 1.5 mM; tube 8: 1.0 mM; tube 9: 2.0 mM.
Tubes 10-16, final Mg++ conc. 1.5 mM; tube 17: 1.0 mM; tube 18: 2.0 mM.
Tubes 19-25, final Mg++ conc. 1.5 mM; tube 26: 1.0 mM; tube 27: 2.0 mM.
Variable is different template DNAs as shown above. All reactions performed at 40C annealing temperature.
More details can be found as needed in Group notebook in molecular lab.
Gels (4/30) from PCR
Lanes 1-9
Lanes 10 - 18
Lanes 19 - 27
2-log ladder molecular weight marker
Wed Group 2:
4 ul of reaction used in TOPO-TA cloning reaction from tubes 3, 11, 20.
Everyone:
Clones obtained, respread and grow as overnights (LB/Spec media).
DNA isolated with Qiaprep, quantified, inserts analyzed by EcoRI digest. Some clone DNAs sent to EtonBioscience for sequencing.
Gels of EcoRI digests of clones
3-CSF1,2
3-DH1, 3-EH1, 20-DH1
11-CML7,8
20-CML2
Could not identify gel for 20-CML3
20-CML9
Clones sent for sequencing:
3-CSF1
3-CSF2
3-DH1
3-EH1
11-CML7
11-CML8
20-CML2
20-CML3
20-CML9
20-DH2
Each student to analyze two different clone sequences (both forward and reverse primer sequences for each clone) using directions provided.
Project Analysis & Report instructions
