Creating DNA Fragments

What are the 3 methods of creating DNA fragments?

• Reverse transcriptase

• Restriction endonuclease

• Gene machine

Advantage of reverse transcriptase method

• mRNA present in cell is intron free

• So cDNA will be intron free

Disadvantage of reverse transcriptase method

• More steps

• So more time consuming

• Technically more difficult

Advantage of restriction endonuclease method

Sticky ends on DNA fragment make it easier to insert to produce recombinant DNA

Disadvantage of restriction endonuclease method

Still contains introns

Advantage of gene machine method

Can design exact DNA fragment you want (e.g: so it has sticky ends)

Disadvantage of restriction endonuclease method

Need to know the sequence of amino acids or bases