1. mRNA is isolated using MACS Technology from total RNA.
2. cDNA is prepared selectively from Reverse Transcription Poly
Chain Reaction (RT-PCR).
- Magnetic-activated cell sorting (MACS) is a
method for separation of various cell populations depending on
their surface antigens (CD molecules). In MACS technology, mRNA
will be isolated .
- Reverse Transcription Poly Chain Reaction : (RT-PCR)
:
Reverse transcription Poly Chain Reaction uses
mRNA rather than DNA as the starting template.
- The enzyme reverse transcriptase uses the mRNA template to
produce a complementary single-stranded DNA strand called cDNA in a
process known as reverse transcription.
- DNA polymerase is used to convert the single-stranded cDNA into
double-stranded DNA.
These DNA molecules will be used as templates for a Poly Chain
reactions.
Reverse transcribed cDNA synthesized under carefully controlled
conditions is a faithful and stable.