What is SacII?
Table of Contents
What is SacII?
The SacII, NEB is: Time-Saver™ qualified for digestion in 5-15 minutes. 100% activity in CutSmart® Buffer (over 210 enzymes are available in the same buffer) simplifying double digests. Supplied with 1 vial of Gel Loading Dye, Purple (6X) Requires two or more sites for cleavage.
What does restriction site mean?
A restriction site is a sequence of approximately 6–8 base pairs of DNA that binds to a given restriction enzyme. These restriction enzymes, of which there are many, have been isolated from bacteria. Their natural function is to inactivate invading viruses by cleaving the viral DNA.
What does a restriction site allow you to do?
They recognize and bind to specific sequences of DNA, called restriction sites. Each restriction enzyme recognizes just one or a few restriction sites. When it finds its target sequence, a restriction enzyme will make a double-stranded cut in the DNA molecule.
What is SacI restriction enzyme?
Thermo Scientific SacI restriction enzyme recognizes GAGCT^C sites and cuts best at 37°C in its own unique buffer. See Reaction Conditions for Restriction Enzymes for a table of enzyme activity, conditions for double digestion, and heat inactivation for this and other restriction enzymes.
How do I choose a restriction site?
When selecting restriction enzymes, you want to choose enzymes that:
- Flank your insert, but do not cut within your insert.
- Are in the desired location in your recipient plasmid (usually in the Multiple Cloning Site (MCS)), but do not cut elsewhere on the plasmid.
What is the bacteria source for SacI?
SacI is a restriction enzyme isolated from the bacterium Streptomyces achromogenes.
How do you know if your restriction digestion was successful?
If the digested product would be visible at a lower coordinate on the gel, it would have made things easy. You can amplify your digested fragment with primer beginning in the flankers region and with only 3-4 bp in the intern 8680 bp region. If you do not get PCR fradments, was the digestion successfully.
What enzyme cuts DNA at specific sites?
restriction endonucleases
Restriction enzymes, also called restriction endonucleases, recognize a specific sequence of nucleotides in double stranded DNA and cut the DNA at a specific location.
How many restriction enzymes are there?
Restriction enzymes recognize short DNA sequences and cleave double-stranded DNA at specific sites within or adjacent to these sequences. Approximately 3,000 restriction enzymes, recognizing over 230 different DNA sequences, have been discovered.
Does SACI have star activity?
Time-Saver™ qualified for digestion in 5-15 minutes. Reduced star activity. Supplied with 1 vial of Gel Loading Dye, Purple (6X)
What are isoschizomers and neoschizomers?
Isoschizomers are restriction enzymes that have the same recognition sequence and cleave the DNA at the same positions, while neoschizomers are restriction enzymes that have the same recognition sequence but cleave DNA at different positions.
