BspQI (Type IIS Restriction Enzyme)

REF: EG23503S

Recognition & Cleavage

Recognition site: 5'-GCTCTTC-3' (non-palindromic)

Cleavage pattern:

5' ... G C T C T T C (N_¹)^▼ ... 3'
3' ... C G A G A A G (N_⁴)_▲ ... 5'

Type: IIS (cuts outside the site) · Common use: Golden Gate assembly

Isoschizomers: SapI, LguI, PciSI (methylation sensitivities may differ)

Specs

Optimal temperature: 50 °C
Heat inactivation: 80 °C · 20 min
Storage: −20 °C

Pack Components

Component	EG23503S
BspQI (10 U/µl)	50 µl
10× Cut Buffer C	1 ml

Recommended Reaction

Buffer: 1× Cut Buffer C (contains recombinant albumin for enzyme stability)
Incubation: 50 °C
Typical duration: 15–60 min (per substrate & load)

Definition of 1 U: amount required to completely digest 1 µg λDNA in 50 µl at 50 °C for 1 h.

Quick Protocol (DNA Digestion)

On ice, assemble 50 µl: ddH₂O up to 50 µl, 5 µl 10× Cut Buffer C, 1 µg DNA, 1 µl BspQI (10 U/µl).
Mix gently, quick-spin.
Incubate at 50 °C for 15–60 min.
(Optional) Inactivate at 80 °C for 20 min or clean up (spin column or phenol/chloroform).

Notes

Keep enzyme volume ≤10% of total (glycerol can promote star activity).
Avoid inhibitors in DNA (phenol, chloroform, ethanol, EDTA, high salt, detergents).
Smaller total volumes amplify inhibition from salts/EDTA/glycerol—scale accordingly.

Quality Control

Prolonged incubation/star activity: 10 U with 1 µg λDNA, 3 h at optimal temp → no nonspecific degradation detected (longer incubation may show star activity).
Ligation & recutting: >95% ligation at 22 °C (T4 DNA ligase); >95% of ligated fragments recut by BspQI.
DNase test: 10 U with 15 ng dsDNA, 16 h at 37 °C → no detectable degradation.

Methylation Effects

Methylase	Effect
Dam	No effect
Dcm	No effect
CpG	No effect
EcoKI	No effect
EcoBI	No effect

Recognition Sites in Common DNAs

DNA	# Sites	DNA	# Sites
λDNA	10	ΦX174	1
pBR322	1	pUC57	1
pUC18/19	1	SV40	0
M13mp18/19	0	Adeno2	7

Write a review

Your Name

Your Review

Note: HTML is not translated!

Rating Bad Good

Captcha

Please complete the captcha validation below

BspQI

Brand: Best Enzymes
Product CAT#: EG23503S
Availability: In Stock

Available Options

Enzymes Handling Fee

Overnight Shipping Handling (+$29.99)

Size

Qty

0 reviews / Write a review