Isolation

Protocol to cryopreserve and isolate nuclei from adipose tissue without dimethyl sulfoxide

M. M. Almeida, Caires, L. C. J., Musso, C. M., Campos, J. M. S., Maranduba, C. M. C., Macedo, G. C., Mendonça, J. P. R. F., and Garcia, R. M. G., Protocol to cryopreserve and isolate nuclei from adipose tissue without dimethyl sulfoxide, vol. 13, pp. 10921-10933, 2014.

Cryopreservation injuries involve nuclear DNA damage. A protocol for cryopreserving and isolating adipocyte nuclei is proposed. Adipose tissue samples were directly analyzed (NoCRYO-0h), or stored at -196°C for 7 days without 10% dimethyl sulfoxide (DMSO) (CRYO-WO-DMSO) or with DMSO (CRYO-W-DMSO). To determine the effect of DMSO on cryopreservation treatment, adipose tissue samples were stored at 4°C for 24 h with 10% DMSO (NoCRYO-W-DMSO-24h) and without (NoCRYO-WO-DMSO-24h). Samples were processed in isolation buffer, and nuclear integrity was measured by flow cytometry.

Isolation and characterization of microsatellite loci from an endangered tree species, Toona ciliata var. pubescens

J. Liu, Sun, Z. - X., Chen, Y. - T., and Jiang, J. - M., Isolation and characterization of microsatellite loci from an endangered tree species, Toona ciliata var. pubescens, vol. 11, pp. 4411-4417, 2012.

Toona ciliata var. pubescens is considered an endangered tree species native to China. In order to help develop a conservation program for this species, we evaluated its genetic diversity and population genetics. We isolated microsatellite DNA loci using streptavidin beads. A genomic library, enriched with microsatellites, was constructed and screened by sequencing. We detected 8 polymorphic microsatellite loci from the tree tissue samples. The population of T. ciliata var.

Molecular cloning, sequence characterization, and gene expression profiling of a novel water buffalo (Bubalus bubalis) gene, AGPAT6

S. Song, Huo, J. L., Li, D. L., Yuan, Y. Y., Yuan, F., and Miao, Y. W., Molecular cloning, sequence characterization, and gene expression profiling of a novel water buffalo (Bubalus bubalis) gene, AGPAT6, vol. 12, pp. 4116-4126, 2013.

Several 1-acylglycerol-3-phosphate-O-acyltransferases (AGPATs) can acylate lysophosphatidic acid to produce phosphatidic acid. Of the eight AGPAT isoforms, AGPAT6 is a crucial enzyme for glycerolipids and triacylglycerol biosynthesis in some mammalian tissues. We amplified and identified the complete coding sequence (CDS) of the water buffalo AGPAT6 gene by using the reverse transcription-polymerase chain reaction, based on the conversed sequence information of the cattle or expressed sequence tags of other Bovidae species.

Isolation and identification of bovine primary hepatocytes

Q. D. Jiang, Li, H. P., Liu, F. J., Wang, X. J., Guo, Y. J., Wang, L. F., Lu, W. F., Li, H. J., Li, X. P., and Yang, G. Y., Isolation and identification of bovine primary hepatocytes, vol. 12, pp. 5186-5194, 2013.

The liver is a unique organ that is endowed with a plethora of specialized functions. Most of its functional traits are controlled by hepatocytes. Primary hepatocytes have been used widely in in vitro models to understand the biological processes occurring in the liver. There are a number of methods used to separate hepatocytes, but the cell activity and purity are much lower in this condition. On the basis of previous research, in this study, the two-step collagenase perfusion technique was used for isolating hepatocytes.

A generic plant RNA isolation method suitable for RNA-Seq and suppression subtractive hybridization

Y. Q. Zhu, Wu, W. J., Xiao, H. W., Chen, H. B., Zheng, Y., Zhang, Y. J., .X.Wang, H., and Huang, L. Q., A generic plant RNA isolation method suitable for RNA-Seq and suppression subtractive hybridization, vol. 12, pp. 5537-5546, 2013.

A recently developed revolutionary approach to transcrip­tomics, RNA-Seq, and suppression subtractive hybridization are power­ful tools for gene expression research. However, currently, the difficulty of isolating high-quality RNAs from plant tissues bearing abundant complex polysaccharides, polyphenolics, and secondary metabolites is a serious problem that not only limits the application of these technologies but also hinders studies dealing with RNA in general.

Development and characterization of 32 microsatellite loci in the giant grouper Epinephelus lanceolatus (Serranidae)

S. Yang, Wang, L., Zhang, Y., Liu, X. C., Lin, H. R., and Meng, Z. N., Development and characterization of 32 microsatellite loci in the giant grouper Epinephelus lanceolatus (Serranidae), vol. 10. pp. 4006-4011, 2011.

An economically important marine fish species, the giant grouper Epinephelus lanceolatus (Serranidae) is widely cultured in Taiwan and costal areas of China. We isolated and characterized 32 polymorphic microsatellite loci from a CA-enriched genomic library of giant grouper. The number of alleles per locus ranged from 3 to 7, with a mean of 4.69. Observed and expected heterozygosities per locus varied from 0.387 to 1.000 and from 0.377 to 0.843, respectively. Six loci significantly deviated from Hardy-Weinberg equilibrium.

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