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Thermostable Endonuclease Q

Catalog # Concentration Size List Price Quantity Your Price
M0701S 50 units $128.00
$115.20
Catalog # Size List Price Your Price
M0701S 50 units $128.00
$115.20
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*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

E1604 Time Temp

  • DNA endonuclease
  • Active on ssDNA and dsDNA
  • Catalyzes the cleavage of a phosphodiester bond 5′ from deoxyxanthosine (dX), deoxyinosine (dI), deoxyuracil (dU), and an abasic (AP) site, creating a nick with 3′-OH and a 5′-phosphate
  • Thermostable
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    What are endonucleases and their applications?

Thermostable Endonuclease Q (EndoQ) is an archaeal DNA endonuclease that cleaves the phosphate backbone of a DNA substrate 5′ to the position of the modified nucleobases of deoxyxanthosine (dX), deoxyinosine (dI), deoxyuracil (dU), and an abasic (AP) site. The result is a 3′-OH and a 5′-phosphate. Thermostable Endonuclease Q generally prefers single-stranded DNA over double-stranded DNA, and exhibits higher activity on dU over dI and dX.

Product Source: An E. coli strain that carries a cloned archaeal Endonuclease Q gene with a C-terminal his-tag.


Figure 1: Relative Activity of Thermostable Endonuclease Q on Various DNA Substrates

Relative Activity of Thermostable Endonuclease Q

Thermostable Endonuclease Q has different specificities for modified nucleotides in ssDNA and dsDNA. Thermostable Endonuclease Q (#M0701) activity was tested on 1 pmol of ssDNA or dsDNA substrates with fluorescently-labeled oligonucleotides containing modified nucleotides of interest (red) for 1 hour in 1x NEBuffer r2.1 at 65°C. Cleaved products were detected by capillary electrophoresis. Thermostable Endonuclease Q prefers ssDNA, and exhibits higher activity on dU over dI and dX.

Figure 2: Temperature Profile of Thermostable Endonuclease Q

Temperature Profile of Thermostable Endonuclease Q
Thermostable Endonuclease Q is active over a wide range of temperatures. Thermostable Endonuclease Q (#M0701, 0.2 U) activity was tested on 1 pmol of a fluorescently-labeled single-stranded oligonucleotide containing a single uracil for 1 hour in 1x NEBuffer r2.1 at various temperatures. Cleaved products were detected by capillary electrophoresis. Thermostable Endonuclease Q is active over a wide range of temperatures (37°C – 90°C), but displays optimal activity between 65°C and 85°C.

Figure 3: Thermostable Endonuclease Q is active in some DNA:RNA contexts

Thermostable Endonuclease Q active in some DNA:RNA
Thermostable Endonuclease Q is active on some DNA:RNA contexts. Thermostable Endonuclease Q (#M0701, 0.2 U) activity was tested on 1 pmol of substrate containing a single deoxyuracil (dU) or ribouracil (rU) within a DNA oligonucleotide (gray) or RNA oligonucleotide (blue) for 1 hour in 1x ThermoPol® Reaction Buffer. Activity was compared with Thermostable USER® III Enzyme (#M5509) for comparison. Cleaved products were detected by capillary electrophoresis. Thermostable Endonuclease Q can cleave at dU and rU located within DNA oligonucleotide strands regardless of presence or type (DNA or RNA) of complement.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Thermostable Endonuclease Q M0701SVIAL -20 1 x 0.05 ml 1,000 units/ml
  NEBuffer™ r2.1 B6002SVIAL -20 1 x 1.25 ml 10 X
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