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NEB Cutter
(Online Restriction Digestion)
1
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
3
4
Custom Digest: Custom digestion involves
digesting With specific or desired enzymes.
5
Selection of Enzyme:
6
Restriction map:
7
Restriction sites:
8
Fragments Length and Position:
9
10
On Gel:
11
12
• 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

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HUMAN EYE By-R.M Class 10 phy best digital notes.pdf
 

Neb cutter

  • 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
  • 9. Fragments Length and Position: 9
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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

Editor's Notes

  1. This digestion involves digesting the sequence with all the available enzymes on NEB.
  2. The required enzymes are selected by viewing their restriction sites and other characteristics from the list of available enzymes,more than one or two enzymes can be selected from the list.