Transformation results: Many tiny colonies for NS3 into HMS174, little tiny colonies for pkd46 into HMS174, no colonies for DH5a.
DH5a cells are probably not competent any more.
Purified colonies by streaking them onto new dishes.

Real-time PCR
Purpose: to quantify amount of transcription in cell
Aim of expt: To find out if csrA, B, C are transcribed at the same level at different phases of E.coli growth and if they are transcribed at the same level for hos and P+ cells
1. Plan where to put each gene in the 96 wells (do in triplicates, technical replicates) (including endogenous control like rpoD, it is supposedly transcribed at a constant level all the time, so comparing to this gene can eliminate errors due to different sample sizes)
2. Label everything. Make sure add the right thing to the right place. Use clean pipettes
3. Dilute primers 10x(?)
4. Total volume in each well= 25ul
5. Add right volume of Mastermix (DNA polymerase, nucleotides, CaCl2), water , and SYBR Green to all primers
6. Add mixture to the right wells
7. Add cDNA to the right wells
8. Change pipette tips after every add. Open a box of new tips, use tips corresponding to the position of the wells. Use tips with the cotton wool. Keep everything clean cos PCR is very sensitive
9. Put in RT-PCR machine, for a 3(?) hours