What is the restriction site for HindIII?

1. The restriction enzyme HindIII cuts DNA at the sequence AAGCTT, and the restriction enzyme HpaII cuts DNA at the sequence CCGG.

How were restriction enzymes used in pGLO?

Restriction digests also form part of the foundation for molecular biology in general. Recombinant DNA molecules (such as the pGLO plasmid) are traditionally made by cutting various desired DNA fragments with restriction enzymes, then ligating (joining) the pieces together.

What is the restriction site of Hind enzyme?

HindIII (pronounced “Hin D Three”) is a type II site-specific deoxyribonuclease restriction enzyme isolated from Haemophilus influenzae that cleaves the DNA palindromic sequence AAGCTT in the presence of the cofactor Mg2+ via hydrolysis.

Where are restriction enzymes located?

bacteria
Restriction enzymes are found in bacteria (and other prokaryotes). They recognize and bind to specific sequences of DNA, called restriction sites. Each restriction enzyme recognizes just one or a few restriction sites.

Does Hind 3 produce sticky ends?

Option B: Hind 3: It is a type 2 restriction endonuclease which gives sticky ends. It is isolated from Haemophilus influenzae.

How do you calculate restriction sites?

First, work out the frequency of occurrence of the restriction site as 1-in-x bases, as explained in the example for the Intermediate level calculation. Then take the size of the DNA in kb (kilobases) and multiply by 1000 to get the size in bases. Divide this by x and round to the nearest whole number.

Why does UV light make pGLO glow?

“The beta-lactamase protein is produced and secreted by bacteria that contain the plasmid. The Green Fluoresce Protein (GFP) is a gene code that is inserted into the pGLO plasmid to make it glow green under UV light with the help of the araC gene.

Why is arabinose needed for pGLO?

Arabinose acts as an allosteric regulator of AraC, changing which DNA sites it binds to and how it forms a dimer. Remember that arabinose is the sugar that gets catabolized by the proteins of the AraBAD operon. When arabinose is added to the environment in which E. coli live, it binds tightly to AraC.

What is the full form of Hind II?

Hind ii. (Science: enzyme molecular biology) first type II restriction endonuclease identified, by Hamilton Smith in 1970. Isolated from haemophilus influenzae, it cleaves the sequence GTPyPuAC between the unspecified pyrimidine and purine generating blunt ends.

How do you know if a site is restrictions?

The option Find Restriction Sites… from the “Tools”→“Cloning” menu or the context menu allows you to find and annotate restriction sites on a nucleotide sequence.

Does Hind 2 produce sticky ends?

Hind II recognizes the sequence GTPy/PuAC and generates fragments with blunt ends (1). Compatible ends Hind II generates fragments with blunt ends and is compatible to any other blunt end. Isoschizomers Hind II is an isoschizomer to Hinc II.

What are the 3 restriction enzymes in pGLO?

The restriction enzymes are: The first three (Hind III, Pvu I, and Ssp I) are 6-base cutters; their restriction sites are 6 nucleotides long. Note that Hind III and Pvu I leave sticky ends, while Ssp I leaves blunt ends: both strands are cut at the same place. Sat I is different from the others.

What are the restriction sites of the pGLO plasmid?

Multiple cloning site — a region containing restriction sites (NdeI, HindIII, EcoRI, etc.), sequences that permit the insertion or deletion of a gene of interest Bacteria transformed with the pGLO plasmid are selected by ampicillin resistance and when induced to express GFP, they glow fluorescent green under UV light!

Which is the best restriction enzyme for HindIII?

HindIII restriction site: DNA Restriction Enzymes from Takara such as HindIII are high-quality: perform restriction enzyme digestion with reliable restriction endonucleases. Cat. # 1060B contains 5 of Cat. # 1060A. Please refer to Cat. # 1060A for complete product documentation and resources. Our products are to be used for Research Use Only.

Can you add restriction enzymes to recipient plasmid?

However, you still need to avoid restriction enzymes that cut within your insert. Adding desired restriction sites to your recipient plasmid : You can modify the MCS of your recipient plasmid using Annealed-oligo Cloning.