Direction: Dr. Singh isolated a new 5-kb gene and wants to determine its sequence using a sequencer that can sequence upto 500 bases in a single reaction. Therefore, she decides to create subclones having suitable-size inserts for sequencing
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Which one of the following will be the most appropriate restriction enzyme for this subcloning?
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- 8-bp cutter
- 6-bp cutter
- 3-bp cutter
- 5-bp cutter
Correct Option: C
Total gene size = 5000bp, Because sequencer can sequence upto 500 bases in a single reaction. Then the fragments are = 5000/500 = 10
Therefore 48 = 6536, 46 = 4096 , 45 = 1024, 43 = 64
Only 3 – bp cutter will generate more than 10 fragments ( 5000/64 = 78). Rest other cutter will generate less than 10 fragments.
