CRSEXP025

=Researchers= Kayla Gogarty Jean-Claude Bradley Dolores Conover (supplied e coli) Kara Spiller (PI in e coli projects) =Objective= To investigate if common H NMR can monitor the growth of e coli. =Procedure= =Results= [|CRSEXP025A-NMR1] First NMR Taken of Control Broth, 25.0C, 16 scans [|CRSEXP025B-NMR1] First NMR Taken of E coli Sample, 25.0C, 16 scans [|CRSEXP025A-NMR2] Second NMR Taken of Control Broth Approx. 19 hours later, 25.0C, 16 scans [|CRSEXP025B-NMR2] Second NMR Taken of E coli Sample Approx. 19 hours later, 25.0C, 16 scans [|CRSEXP025A-NMR3] Third NMR Taken of E coli Sample Approx. 5 minutes later, 25.0C, 128 scans [|CRSEXP025C-NMR1] First NMR Taken of CDCl3 Only, 25.0C, 16 scans 1.The main difference in the addition of the "e. Coli" was the appearance of a large amount of glycerol (3.5-4.0 ppm) 2. No obvious changes were noted after 24h for the e. Coli sample or the broth control 3. The water peak was clearly shifted from the usual 4.80 pppm to 4.77 ppm =Discussion= 1. No aromatic peaks were detected in any sample - this is consistent with low values of aromatic amino acids in the broth (MillersLB Broth composition ) 2. A large amount of glycerol was present after the addition of the e coli seed, which was used as a cryogenation preservative (see spectrum section ) 3. The shift in the water peak by 0.03 ppm is significant and is probably reflected by the higher ionic strength from adding NaCl but remains to be tested. =Conclusion= A first attempt shows that there is enough concentration of materials to detect significant changes of composition but none was found after 24h for this particular run to be able to estimate e coli population growth or chemistry changes. It has been noted that bacteria selectively metabolize different sugars. This type of experiment should work by simple H NMR. =Log= Scan Parameters: 16 scans, 1s relaxation delay, 45degree pulse angle Scan Parameters: 16 scans, 1s relaxation delay, 45degree pulse angle Scan Parameters: 16 scans, 1s relaxation delay, 45degree pulse angle Scan Parameters: 128 scans, 1s relaxation delay, 45degree pulse angle Scan Parameters: 16 scans, 1s relaxation delay, 45degree pulse angle Scan Parameters: 16 scans, 1s relaxation delay, 45degree pulse angle
 * [This is needed in more detail from the e coli providers JCB]**
 * 2013-09-25**
 * 5:23**-Control Broth scanned in NMR at 25.0C
 * 5:46**-E coli sample in Broth scanned in NMR at 25.0C
 * 2013-09-26**
 * 12:08**-E coli sample scanned in NMR at 25.0C
 * 12:13**-E coli sample scanned in NMR at 25.0C
 * 12:25**-Control Broth scanned in NMR at 25.0C
 * 12:29**-Outside NMR Tube with CDCl3 scanned at 25.0C to determine if acetone peak in NMR from the outside tube