obtaining DNA fragments - reverse transcriptase

Question 1

explain how you obtain DNA fragments by using reverse transcriptase

Answer 1

• mRNA is found in cells that produce the required protein
• reverse transcriptase makes CDNA
• DNA helicase breaks hydrogen bonds causes strands to separate
• DNA polymerase joins free nucleotides by forming phosphodiester bonds

Question 2

explain how complementary DNA is made using reverse transcriptase

Answer 2

1. strand of mRNA from cells that produce the desired protein
2. mRNA acts as a template for the production of single stranded CDNA using reverse transcriptase
3. CDNA is isolated by hydrolysis of mRNA with DNA helicase
4. double stranded DNA is formed on template of CDNA using DNA polymerase
5. this is the gene for a protein