Prodcuing DNA Fragments Flashcards
What are restriction enzymes
Cuts DNA at specific base sequence called recognise sites
What is the recognition site specific to
Restriction enzyme
Why is recognitions site a Palindrome
Base sequence is the same as its reverse complement of bases
Why are sticky ends “sticky”
Form hydrogen bonds with the complementary bases of the sticky ends of the fragments of other DNA molecules cut by same restriction enzyme.
Blunt ends
Restriction enzyme cuts a straight line
How can fragment be separated
According to size via gel electrophoresis
How can DNA fragments containg gene of interest be identified
Using a gene probe
How is a single stranded cDNA formed
- Through reverse transcription of mRNA
- produces complementary single stranded cDNA
- catalysed wit reverse transcriptase
Does cDNA contain introns
No
Gene machine
Summarise the process of using reverse transcriptase to produce DNA fragments
- mRNA complementary to the target gene is used as a template
- its mixed with free nucleotides which match up to base pairs
- RT which forms the sugar phosphate backbone to relate cDNA
Summarise the recess of using enzymes to produce DNA fragments
- Registration enzymes (endonucleases) cut DNA at specific sequences
- different RE cut at different points but one RE will always cut at the same sequence
- using particular RE allows you to cut out retain gene of interest
