Question

In: Biology

You have total RNA extract from plant. How can you make cDNA only from mRNA selectively?

You have total RNA extract from plant. How can you make cDNA only from mRNA selectively?

Solutions

Expert Solution

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.

  1. 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.
  2. 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.


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