AluI NEB#R0137

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	After digestion with HaeIII and AluI (NEB, New England Biolabs, USA) DNA was treated with MBN nuclease (NEB) and FastAP alkaline phosphatase (Fermentas, Lithuania).

Protocol tips
After digestion with HaeIII and AluI (NEB, New England Biolabs, USA) DNA was treated with MBN nuclease (NEB) and FastAP alkaline phosphatase (Fermentas, Lithuania).

Publication protocol

Total genomic DNA for this study was extracted from a liver tissue of the edible dormouse specimen died during hibernation in the Zoological Garden in Poznan, 1997. After digestion with HaeIII and AluI (NEB, New England Biolabs, USA) DNA was treated with MBN nuclease (NEB) and FastAP alkaline phosphatase (Fermentas, Lithuania). Then, column-purified DNA fragments were ligated to the double-stranded SNX linkers as described by Hamilton et al. (1999). Ligation success was monitored by PCR with SNX forward primer. Ligation mixture was denatured at 95 °C for 5 min and hybridized to 5′-biotinylated (CA)12 and (CT)12 oligonucleotides at 60 °C for 1 min, and then incubated in rotation at 60 °C for 15 min with streptavidin-coated paramagnetic beads (Dynabeads, Life Technologies, USA). Repeat-enriched fragments were PCR amplified using SNX forward primer and cloned into pCR4-TOPO vector using TOPO TA Cloning Kit (Life Technologies). After screening, 96 positive clones were sequenced on an ABI 3130XL DNA analyzer (Applied Biosystems). After excluding clones with <15 repeats, we found 27 non-redundant microsatellite sequences flanked by regions sufficient to design primers. However, among primer sets developed for these loci, only 14 gave specific PCR products and/or were polymorphic in 15 tested specimens representing Croatian and Polish populations

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Restriction Enzymes AluI using AluI NEB#R0137 from New England BioLabs.

Manufacturer protocol

Download the product protocol from New England BioLabs for AluI NEB#R0137 below.

Videos

Check out videos that might be relevant for performing Restriction Enzymes AluI using AluI NEB#R0137 from New England BioLabs. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.