FastDigest MspI

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	15 µl of the resulting PCR product was digested with 2 µl of MspI enzyme (fast digest) and 3 µl enzyme buffer and then completed with nuclease-free water to a final volume of 30 µl, and incubated at 37°C for 30 min	Digested PCR products were electrophoresed using 3% agarose gel to distinguish between the 175 bp band produced by the digestion of the A allele and the 141 bp band produced by the digestion of the G allele. Heterozygous state yielded both 141 bp and 175 bp bands as previously described by Le Marchand et al. [19]

Protocol tips
15 µl of the resulting PCR product was digested with 2 µl of MspI enzyme (fast digest) and 3 µl enzyme buffer and then completed with nuclease-free water to a final volume of 30 µl, and incubated at 37°C for 30 min
Downstream tips
Digested PCR products were electrophoresed using 3% agarose gel to distinguish between the 175 bp band produced by the digestion of the A allele and the 141 bp band produced by the digestion of the G allele. Heterozygous state yielded both 141 bp and 175 bp bands as previously described by Le Marchand et al. [19]

Publication protocol

"Restriction fragment length polymorphism for the PCR
product for detection of CCND1 G870A polymorphism
The resulting 212 base pair (bp) PCR product was digested with the restriction enzyme Moraxella species
(MspI) (Fermentas ) as follows:
15 µl of the resulting PCR product was digested
with 2 µl of MspI enzyme (fast digest) and 3 µl enzyme buffer and then completed with nuclease-free
water to a final volume of 30 µl, and incubated at
37°C for 30 min.
Digested PCR products were electrophoresed using
3% agarose gel to distinguish between the 175 bp band
produced by the digestion of the A allele and the 141 bp
band produced by the digestion of the G allele. Heterozygous state yielded both 141 bp and 175 bp bands as
previously described by Le Marchand et al. [19]."

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Manufacturer protocol

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