Neb cutter

A Software used for visualizing & cutting our DNA of interest with Restriction enzymes. see how this software works?

1. NEB Cutter (Online Restriction Digestion) 1

2. Introduction: NEBcutter accepts an input sequence, which can be pasted in, picked up from a local file or retrieved from NCBI as a GenBank file via its accession number. Various options are available to select the size of ORFs to be displayed and the set of restriction enzymes to be used. The program calculates the positions of all restriction enzyme sites noting those that might potentially be blocked by overlapping methylation and finds the ORFs in the sequence. It then displays a schematic diagram of the sequence, the long ORFs, based on the rules described in the Methods and all restriction enzymes that cut it just once. 2

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5. Custom Digest: Custom digestion involves digesting With specific or desired enzymes. 5

6. Selection of Enzyme: 6

7. Restriction map: 7

8. Restriction sites: 8

9. Fragments Length and Position: 9

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11. On Gel: 11

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13. How to calculate the size of fragments produced by restriction digestion? Subtracting the initial base number from ending base number and then Adding one. For example: BamHI Finds its restriction sites at 5506 bp and 22346bp so the size of its fragment will be 16840bp as explained below: BamHI-BamHI    5506-22346 22346-5506=16840 16840+1=16841bp 13