BseLI (BslI) (10 U/µL)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Restriction analysis was performed with 12 µL of purified PCR product and 1 U of the respective enzyme [MwoI (neoschizomer of HpyF10VI) (Fermentas ER1731/300U) or BslI (isoschizomer of BseLI-BsiYI) (Fermentas ER1202 /2500U)] at 37 °C for 2 h for MwoI and at 55 °C for 45 min for BslI (13).

Protocol tips
Restriction analysis was performed with 12 µL of purified PCR product and 1 U of the respective enzyme [MwoI (neoschizomer of HpyF10VI) (Fermentas ER1731/300U) or BslI (isoschizomer of BseLI-BsiYI) (Fermentas ER1202 /2500U)] at 37 °C for 2 h for MwoI and at 55 °C for 45 min for BslI (13).

Publication protocol

"PCR products were purified using the PCR Clean-Up
Kit (GeneMark DP04) according to the instructions of
the manufacturer. Then REA was applied to all strains.
All strains were digested with REA by using MwoI (5’...G
C N N N N N↓N N G C...3’). Because C. tropicalis and
C. parapsilosis strains could not be discriminated after
digestion with MwoI, further REA was performed by
using the BslI (5’...C C N N N N N↓N N G G...3’) enzyme.
Restriction analysis was performed with 12 µL of
purified PCR product and 1 U of the respective enzyme
[MwoI (neoschizomer of HpyF10VI) (Fermentas ER1731
/300U) or BslI (isoschizomer of BseLI-BsiYI) (Fermentas
ER1202 /2500U)] at 37 °C for 2 h for MwoI and at 55 °C
for 45 min for BslI (13).
Restriction fragments were separated by
electrophoresis in 2% agarose gel at 120 V for 40 min.
An O’RangeRuler 50-bp DNA Ladder (MBI Fermentas
SM0613) was used as a DNA marker. The gels were stained
with ethidium bromide and visualized with Syngene
GeneSnap (Synoptics Ltd., USA)."

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for BseLI (BslI) (10 U/µL) below.

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