5'-C^T C G A G-3'
3'-G A G C T^C-5'
Source:
Xanthomonas holcicola
Form:
Supplied as a liquid in 10mM Tris-HCl (pH 7.4 at 25 degrees C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.
R1625 (double digests): Supplied as a liquid in 33mM Tris-acetate, (pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. Incubate at 37 degrees C
R1625-04 (for 100% digestion): Supplied as a liquid in 10mM Tris-HCl, pH 8.5, 10mM MgCl2, 100mM KCl and 0.1mg/ml BSA. Incubate at 37 degrees C.
Unit Definition:
One unit is defined as the amount of XhoI required to digest 1ug of lambda DNA in 1 hour at 37 degrees C in 50ul of assay buffer.
Protocol for Digestion:
Add:
Nuclease free water: 16ul
R1625-04: 2ul
DNA (0.5-1ug/ml): 1ul
X1030: 0.5-2ul
Mix gently and spin down for a few seconds. Incubate at 37C for 1-16 hours.
Protocol for Digestion Directly after Amplification:
Add:
PCR Reaction Mixture: 10ul (~0.1-0.5ug of DNA)
Nuclease free water: 18ul
R1625-04: 2ul
X1030: 1-2ul
Mix gently and spin down for a few seconds.
Incubate at 37C for 1-16 hours.
Storage and Stability:
May be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
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