BIOTECHNOLOGY ENTREPRENEURSHIP

SPECIAL TRAINING (BeST)

PROGRAMME
INTERNSHIP

MONTHLY REPORT
23 OCTOBER 2012- 23 NOVEMBER 2012

Written by:
CHERYL YEAP SOO YEAN

MONTHLY REPORT
AREAS OF INTERNSHIP

: AIMST UNIVERSITY

SUPERVISOR

: DR. LEE SU YIN

Week 1:
1.) I am continued with the incubation of Pseudomonas aeruginosa in minimal
media supplemented with polytethylene powder. From the culture, I did serial
dilution by serial dilute 100l culture into 900l fresh minimal media until the
dilution factor of 10-6. Meanwhile, 50l from each dilution factor of 10-5 and 106
will be platted on LB agar for bacterial colonies observation.
2.) I am also continued doing RNA extraction from the incubated minimal media.

MONTHLY REPORT
AREAS OF INTERNSHIP

: AIMST UNIVERSITY

SUPERVISOR

: DR. LEE SU YIN

Week 2:
1.) Continue from week 1s work, I did analysis of the bacterial count and drew a
line graph representing the colony forming unit/ml Vs time.

Luminescence sensitivity value, amol

ATP/well

2.) I did an experiment to test the efficiency of chemiluminescence substrate

catlayses by the enzyme horse radish peroxidase with 2 detection methods.
One is using UV tranilluminator to view the light signal emitted from the
reaction. Another one is using ELISA plate reader to quantify the
chemiluminescence signal, and check the half life of the chemiluminescence
substrate. A graph of the chemiliminescence efficiency was generated.

5000000

Luminescence value Vs Time

4500000
4000000
3500000
3000000

1:1 dilution

2500000

No dilution

2000000

Blank

1500000
1000000
500000
0
-500000 0

10

20

30

40

time (minutes)

3.) I did primers designing for Vibrio cholera for the molecular detection of Vibrio
cholera by PCR.
4.) I am also prepared Luria Bertani broth by mixing 10g of tryptone, 10g NaCl, 5g
Yeast Extract and mix into 1L of distilled water. Adjust the pH to 7.2 by NaOH.
Aliquot 10ml into each individual universal bottle. Bring to autoclave.

MONTHLY REPORT
AREAS OF INTERNSHIP

: AIMST UNIVERSITY

SUPERVISOR

: DR. LEE SU YIN

Week 3:
1.) Luria Bertani broth is prepared by mixing 10g of tryptone, 5g of yeast extract,
10g of sodium chloride into 1000ml of distilled water. Adjust the pH to 7.2.
Aliquot 10ml of Luria Bertani broth into universal bottle respectively.
2.) Luria Bertani agar was prepared by mixing 20g of LB powder and 15g of agar
powder into 1000ml of distilled water. Adjust the pH to 7.2. Bring to autoclave.
3.) I prepared bacterial lysate for Polymerase Chain Reaction and Sau-02 gene
were be used as primers for the amplification of MRSA and S. aureus. Different
strains of S. aureus were used for the analytical specificity testing of MRSA
using Sau-02 gene.
4.) I am also prepared glycerol stock for different strains of Salmonella typhi.

MONTHLY REPORT
AREAS OF INTERNSHIP

: AIMST UNIVERSITY

SUPERVISOR

: DR. LEE SU YIN

Week 4:
1.) I am continued doing the specificity test of MRSA where I need to optimize the PCR
reaction in order to get a nice results as sometimes the PCR machines are not
functioning properly. Some wells in the PCR machine does not heated up properly.
This will interfere the denaturation, annealing, and extension processes.

2.) mec-A primers is also used for the molecular detection of MRSA.
3.) Gradient PCR is also done to check the best annealing temperature for mec-A
primers. In conclusion, 54oC is the optimum temperature for mec-A primers.

54oC

4.) Mec-A gene should amplify only in MRSA, not both MRSA and S. aureus. Unlike
Sau-02 gene, Sau-02 gene is present in both MRSA and S. aureus. Hence, Sau02 gene can be used to detect both the bacteria at the same time.
5.) PCR for mec-A primers is still under optimization. More experiments need to
done in order to get a good results of PCR.

SUMMARY/REMARKS/FEEDBACK :
1. Supervisor:

[The trainer(s) performance knowledge, presentation, etc. and facilities

provided]

2. Internship Areas:

[The content of the subject Subject coverage]

3. Areas of Improvement/ Recommendation:

[Based on your evaluation during the session, kindly express your ideas to contribute
further to this module]