Professional Documents
Culture Documents
Intro Lab
Thalassemia
191211
Intro Lab Thalassemia
Bu Lely
Introduction Lab Act FBS 4
Thalassemia
Heterogeneous group of inherited disorders hemoglobin synthesis
Characterized by the absence or reduced output of one or more of
globin chains of hemoglobin
Hemoglobin (Hb)
Normal Adult :
Hb A (22) 97 %
Hb A2 (22) 1-2 %
Hb F (22) 0,5 1 %
Pathophysiology
Deficient hemoglobin (Hb) synthesis resulting from deficient
production of one or another of the polypeptide chains of globulin
Red cell membran demage
Shortened red cell survival
Page | 2
Animal Cell
Isolation of DNA
Page | 3
Distruption of the cell membrane
Disosiasi of the protein DNA complexes
PCR
Page | 4
Specific DAN sequence amplification techique to enable further
DNA analysis
Principle : - DNA replication proses
DNA template
Primer (20-25 nucleotida)
Polymerase Enzyme (Taq Polimerase)
Substrate (dNTP
Difference PCR : DNA unwind through
temperature change (high temperature)
Replication : DNA unwind by enzyme
Denaturation
Denaturation at 94C :
Extension
extension at 72C :
Page | 5
Page | 6
1.8E+09
1.6E+09
1.4E+09
1.2E+09
8.0E+08
6.0E+08
4.0E+08
2.0E+08
0.0E+00
0 5 10 15 20 25 30 35
PCR cycle
10
7
DNA copy number (log)
0
0 5 10 15 20 25 30 35
PCR cycle
Electrophoresis DNA Resolution of DNA fragments On agarose gel
Page | 7