ASMA Training _ Introduction

Main Projects - ASMA Training

The training provides various molecular assays used in diagnosis and treatment of chronic myeloid leukemia.

The molecular biology technologies involved in molecular assays are as follow:

- Real-time quantitative PCR (RQ-PCR): to measure BCR-ABL transcript level

- Multiplex PCR: to identify the type of BCR-ABL transcripts

- Direct sequencing & Allele specific oligonucleotide PCR (ASO-PCR):

to detect mutations in Bcr-Abl kinase domain

- Molecular cloning: to prepare reference and target plasmid standards for absolute quantification in RQ-PCR.

- Other technologies: White blood cell isolation, RNA extraction & quality check,

DNA extraction, reverse transcription, gel electrophoresis etc.

According to training duration, three different courses will be provided.

- Yellow course (1 week) covers RQ-PCR

- Blue course (2 weeks) covers RQ-PCR and mutation analysis

- Red course (3 weeks) covers RQ-PCR, mutation analysis and molecular cloning

- Black coure (4 weeks) covers RQ-PCR, mutation analysis, molecular cloning and pharmacokinetics.

Training Program
Assay	Technology	Yellow 1 week	Blue 2 weeks	Red 3 weeks	Black 4 weeks
BCR-ABL transcript type & transcript level analysis	White blood cell purification	O	O	O	O
	RNA isolation & quality check	O	O	O	O
	First strand cDNA synthesis	O	O	O	O
	Multiplex PCR	O	O	O	O
	RQ-PCR	O	O	O	O
	Real time PCR data analysis	O	O	O	O
Mutation analysis in Bcr-Abl kinase domain	Long template PCR		O	O	O
	Nested PCR		O	O	O
	Big Dye termination PCR		O	O	O
	DNA sequencing		O	O	O
	DNA sequencing data analysis		O	O	O
	DNA extraction & ASO-PCR		O	O	O
Molecular cloning	Plasmid ligation			O	O
	Transformation of competent bacteria			O	O
	Colony formation			O	O
	Colony PCR			O	O
	Recombinant plasmid preparation			O	O
Pharmaco-kinetics	HPLC				O
Pharmaco-kinetics	Tandem Mass Spectrometry				O

Training for each assay consists of the following components;

- Introduction of background knowledge and technical information by training staff

- Full demonstration of every step of each assay by qualified demonstrator.

- Practical performance of the demonstrated steps by trainee under the supervision of demonstrator.

- Actual performance of whole assay by trainee