Restriction sites should be common so that gene of interest will be The most important thing is to find the restriction Click ok and it will beД¬hoose DNA select there will be option of this new New DNA file in SnapGene and name this file.

Sequence will be copied with numbers and paste in a Both score are available here.ДЄfter adaptation our sequence will be purified and protein Next there will be improved DNA, underlines show that GC content is supposed to be in between 40 to Important values CAI value that is also shown in graph. Select ECOR1ДЄfter submitting, details will be shown here. Option then organism name means with which organism we want to check theДЄll additional options will be selected. Now the second thing is Gene of interest.ДЇor preparation of gene of interest, java codonД¬opy any gene of interest and then paste that gene onДЄll the options will be selected including the DNA Pet vector is available on this page with all the Now on homepage of SnapGene there is a option of Import this vector and new window will be open there, all the details will be For example write pet 28 in search and the vector will be selected and Here, snapgene online sequences are available. Other option is select files and click import, online databases will be shown Vector can be directly downloaded from google and the Homepage of SnapGene cloning will be open. The tool used for this purpose is SnapGene.ДЇirst click action then there will be other options.ДЇirst one is restriction and insertion cloning, then click on insert fragment. How cloning is performed using restriction enzyme and GENE CLONING, RESTRICTION AND INSERTIONAL CLONING: The above link is provided to perform this. There will be specifically restriction sites of only those enzymes that were selected.ДЄll the fragments of that specific enzyme will be shown here. If we specifically want to check the action of enzymes on our sequence we will select two enzymes from all enzyme option on our home page and then instead of selecting map sites we will have to select Virtual digest. If we talk about specific we can select one or two enzymes that are specific in cloning (on our input page )or transformation and then there digestion will be checked. And in the end there will be cut positions. Site length and overhang information will be shown here. Map site: Map site is used for general means we can check restriction sites by selecting all enzymes.ДЇirst of all there are name of enzymes then the sequence. In Sequence info, we will paste the sequence and write the name of sequenceВ also. Prototype only (select this option ) means original enzymes will be used. While new england bio lab will only add those that have restriction endonucleases.ДҐХњ overhang, 3Хњ overhang and blunt end will be selected. Next there are two options All commercial and NEB, we will select All commercial that means any enzyme that is available on any of database that will be included. In include option there are options for different enzymes that we want to select. Minimum site length should be of 5 bases. Overhang means either there is 5 end or 3end or blunt end, we choose 5 and 3 end.ДЇor Maximum cut there is no restriction. Linear sequence will be selected for DNA and for plasmid: circular option will be selected.ДЇrequency option is already selected by default. To find restriction sites from a DNA or a gene the most important tool that is used is Restriction Mapper.ДЄn online available tool that is required in order to perform cloningДЇirst of all we select the basic options in home page. RESTRICTION ENZYME DIGESTION FOR CLONONIG:

Snapgene viewer restriction digest