So happy I could die!!!!
- Today, Melvin started doing the LRE transformation described last Thursday (as ligations were left the whole weekend).
- 1: LRE (XBA I/PST I) with J23100 (ampicilin).
- 2: LRE (ECO RI/PST I) with plasmid 17 labeled as LRE + p17 Mar and date (chloramphenicol).
- 3: Control of J23100 (ampicilin).
- 4: Control of plasmid 17 (chloramphenicol).
- 5: Control of transformation (BBa_I51020) (ampicilin).
- 6: Control of competent cells.
Primer's for Click Beetle Luciferase
- As Dr. Chris Wood gave us two vectors containing luciferases from click beetle (Chroma-Luc™ Reporter Vectors CBRluc and CBG99luc, Promega Corporation), but which don't have the registry's standard, I did primers to add the preffix and RBS, and the suffix. This will help me to work with them.
GAA TTC GCG GCC GCT TCT AGA GAT TAA AGA GGA GAA AAT GGT GAA GCG TGA GAA AAA TG
Where bold letters indicate the preffix, italics indicate a strong RBS, and normal letters indicate the beginning of the sequence.
CTG CAG CGG CCG CTA CTA GTA TTA TTA ACC GCC GGC CTT CTC
Where bold letters indicate the suffix and normal letters indicate the beginning of the sequence.
- To continue proving the luciferase, I made a restriction for a total of 30μl as follows for both luciferases.
-H2O -------> 8μl
-Buffer 2 --> 3μl
-BSA ------> 1μl
-DNA -------> 15μl
-SPE I -----> 1.5μl
-PST I ------> 1.5μl