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1. PCR-BASED EVALUATION OF THE BETA THALASSEMIA MUTATIONS
IN A PATIENT BLOOD-DERIVED GENOMIC DNA SAMPLE
SUBMITTED BY,
P.Praveena 2 M.sc-
MICROBIOLOGY

2. CONTENTS
• Introduction
• Symptoms
• Diagnosis
• Treatment
• Aim
• Objective
• Materials
• Procedure
• Result
• Conclusion

3. INTRODUTION
• Beta Thalassemia is a genetic blood disorder caused by reduced
production of beta-globin chains.
• It is mainlytwo types. They are thalassemia major and
thalassemiaminor
• Major is the severe form bcz production of beta-globinchains absent.
• Minor is middle form reduction in beta-globin production is less.

4. SYMPTOMS
• Beta thalassemia may lead to anemia
• Bone deformities
• Fatigue
• Cardiac complications
• Growth and developmentissues
• Weakness
• Jaundice

5. DIAGNOSIS
• Structure Hb testing
• CBC(complete blood test)
• Genetic testing
• Haemoglobin electrophoresis
• Iron studies
• Medical history.

6. TREATMENT
• Bone marrow transplantation
• Gene therapy
• Blood transfusions
• Chelation therapy
• Folic acid supplements

7. AIM&OBJECTIVES
• AIM:
• The project work is focused beta Thalassemia mutations in a patient blood derived genomic DNA sample
by using PCR and Gel electrophoresis.
• Objective:
• The main objective is to achieve accurate and early diagnosis for effective management and treatment
of the condition.

8. MATERIALS
• 2x PCR Taq polymerase
• Primer mixture of internal control
• Patient genomic DNA
• Molecularbiology grade water
• Internal control primers
• TAE buffer
• Agarose powder
• SYBR- safe

9. PROCEDURE
• Master mix preparation:
• Take PCR tubes and add 25 microlitersof 2X PCR Taq
polymerase into each tube
• Add 2 microliters of primers mixture of internal control
• After add 2 microliters of genomic DNA
• Add 19 microliters of MBGW then add 2 microliters of primers.
• Place tubes in thermal cycler and set temperatures.

11. PROCEDURE
• Agarose gel preparation:
• prepare buffer by adding 1.6ml of TAE buffer in 196 ml of di-water.
• prepare 2% agarose gel and weigh 1.6 ml of agarose powder and dissolve in 80ml of
di-water.
• Agarose powder mix in conical flask and keep this in oven and cool content.
• And 8 microliters of SYBR-safe. *pour agarose into a gel casting tray.
• place combs over it and leave until becomes solidified.
• collect DNA samples and add loadingdye.
• Remove combs and remove gel from casting tray and transfer into electrophoresis
chamber.
• pour 350ml of buffer over gel and load DNa ladder to 1st well and load the samples
into another well.
• observe bands by using U.V transilluminator.

12. RESULT
• Mutationsare observed in 214 base pair length.
Based on DNA ladder bands at 861 base pair
Length not effected with beta-thalassemia.
• Based on beta, beta° type consider it as a beta
minor thalassemia case.

13. CONCLUSION
• In conclusion, PCR-based evaluation of beta-thalassemia mutationsin a patient's genomic DNA sample is a
vital diagnostictechnique that enables the identification ofgenetic variants associated with this disorder. The
results obtained from this analysis play a crucial role in diagnosing beta-thalassemia and guiding clinical
management decisions.

14. Thank you