- inverse PCR uses for amplifying the DNA sequence that is still unknown

- RFLP (restriction fragment length polymorphism)

- polymorphism = many-shaped/differences among people in their DNA seq at sites recognized by
restriction enzymes
- RT-PCR (reverse transcriptase)
- more mRNA, more expression (e.g., if gene RED has less mRNA than gene BLUE and the inheritors will
become more blue, bcs blue gene carried by more proteins)
- cDNA is DNA that has RNA converted back into DNA copy
- mRNA : 5’-AUG GUC-3’
- cDNA : 3’-TAC CAG-5’ (complementary to mRNA)
- purpose of RT-PCR : to understand the mRNA, to make intron free DNA to manipulate
- in the electrophoresis, DNA is negative and should be run into positive (red wire) and opposite with the
positive (black wire)
- the more the RNA, the thicker the band it will be
- up regulated : more expression, down regulated : less expressed
- Differential Display PCR : combination of RAPD & RT-PCR
- oligo(dT) primers is used to amplified sequence
- the random primer, bcs of it is the mixture primer it will bind in somewhere in the mRNA
- compare the bands (e.g., smoker and non-smoker genes) and there will be some genes missing in the
non-smoker genes, which are appear in the smoker genes (bands). the genes for the smoker is up
regulating, so the bands could be cut-off and use the RT-PCR or qualitative PCR to know the sequence.
- RACE is used to rescue the lost ends of clone genes
- anchor sequence
- hybrid gene : bind overlap primers
- directed mutagenesis
- mismatch in the primer will make a bump
- bp = base pairs
- site directed mutations = mismatch in the primer
- AUC CGU GCU -> ile arg ala
- AUC ACG UGC U -> ile thr cys
- a one insertion could make the whole codons changed