Pellet Pestle Homogenization of Agarose Gel Slices at 45°C for Deoxyribonucleic Acid Extraction

Kurien, Biji T.; Kaufman, Kenneth M.; Harley, JB; Scofield, R. Hal

doi:10.1006/abio.2001.5299

Cited by 14 publications

(11 citation statements)

References 16 publications

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“…This is attractive if one can be satisfied with a yield of 50%. In our laboratory, we obviously prefer to use our own extraction procedure (67) which is very simple to use and provides a consistently good-quality DNA in very good amounts.…”

Section: Discussionmentioning

confidence: 99%

Extraction of Nucleic Acid Fragments from Gels

Kurien

Scofield

2002

Analytical Biochemistry

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Section: Discussionmentioning

confidence: 99%

Extraction of Nucleic Acid Fragments from Gels

Kurien

Scofield

2002

Analytical Biochemistry

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“…Amplification products were separated by electrophoresis on 2% agarose gels. PCR products of the desired size were extracted from the gels by pestle homogenization at 45-C (Kurien et al 2001). PCR products were blunt-end ligated into pUC18 plasmids (Sambrook & Russell 2001).…”

Section: Amplification Cloning and Sequencing Of Te Fragmentsmentioning

confidence: 99%

Ribosomal DNA locus evolution in Nemesia: transposition rather than structural rearrangement as the key mechanism?

2006

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We investigated chromosome evolution in Nemesia using fluorescent in-situ hybridization (FISH) to identify the locations of 5S and 45S (18-26S) ribosomal genes. Although there was conservation between Nemesia species in chromosome number, size and centromere position, there was large variation in both number and position of ribosomal genes in different Nemesia species (21 different arrangements of 45S and 5S rRNA genes were observed in the 29 Nemesia taxa studied). Nemesia species contained between one and three pairs of 5S arrays and between two and four pairs of 45S arrays. These were either sub-terminally or interstitially located and 45S and 5S arrays were often located on the same chromosome pair. Comparison of the positions of rDNA arrays with meiotic chromosome behaviour in interspecific hybrids of Nemesia suggests that some of the changes in the positions of rDNA have not affected the surrounding chromosome regions, indicating that rDNA has changed position by transposition. Chromosome evolution is frequently thought to occur via structural rearrangements such as inversions and translocations. We suggest that, in Nemesia, transposition of rDNA genes may be equally if not more important in chromosome evolution.

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“…1d and Supplementary Table 1) 9 . Purification and analysis of each structure by TEM and agarose-gel electrophoresis indicated enrichment of the properly folded structures and yields of 70%, 50% and 45% for the twelve-helix bundle, ring and tensegrity structure, respectively—values up to four-fold greater than achieved by the pellet-pestle homogenization method 7 .…”

mentioning

confidence: 88%

“…3). ImageJ analysis of the purified six-helix bundles indicated 71±3% of the well-folded structure could be recovered from the agarose matrix versus 15±5% by the pellet-pestle homogenization method 7 . Our analysis by negative-stain transmission electron microscopy (TEM) also indicated a strong enrichment of the properly folded structures.…”

mentioning

confidence: 99%