A rapid and effective method for silver staining of PCR products separated in polyacrylamide gels

Liang, Qingzhi; Wen, Dingqing; Xie, Jianhua; Liu, Liqin; Wei, Yongzan; Wang, Yicheng

doi:10.1002/elps.201400182

Cited by 20 publications

(20 citation statements)

References 12 publications

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“…Silver staining of DNA in polyacrylamide gels has the highest sensitivity compared with other staining methods , and has been widely used to detect DNA fragments such as SSR and other types of DNA fragments as molecular markers . Like many other biological laboratory techniques, silver staining of gels has been steadily improved since its first report as a fragment visualization technique in 1979 .…”

Section: Reagents Process Time and Minimum Amount Of Dna Used In DImentioning

confidence: 99%

“…However, most of these updated protocols still have some drawbacks because they involve high technical demand and time‐consuming processes in fixation and mounting steps . These drawbacks make application of the silver staining more difficult and resource consuming . Further optimization of the protocols by reducing cost and improving efficiency of fragment detection will facilitate routine application of the technique in biological research, especially in large‐scale genotyping.…”

Section: Reagents Process Time and Minimum Amount Of Dna Used In DImentioning

confidence: 99%

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Development of a simple and effective silver staining protocol for detection of DNA fragments

Liu

Ayalew

et al. 2017

Electrophoresis

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Silver staining is one of the widely used methods for DNA fragment detection in biological research. Silver staining protocols have been steadily optimized to improve detection efficiency. This research reports a continuous effort to simplify the existing silver staining protocols, lower experiment cost, and improve DNA detection sensitivity and image clarity. The new method only requires three reagents (silver nitrate, sodium hydroxide, and formaldehyde) and 6-7 min with high detection sensitivity to visualize as low as 14.6 pg (3.3 pg/mm ) of DNA in a non-denaturing polyacrylamide gel. In comparison to previous reported protocols, the new one has the highest resolution, is the easiest to operate, takes the shortest time, and uses the fewest chemical reagents. Therefore, the new method can be used for quick generation of high quality molecular marker data in genetic analysis.

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Section: Reagents Process Time and Minimum Amount Of Dna Used In DImentioning

confidence: 99%

Section: Reagents Process Time and Minimum Amount Of Dna Used In DImentioning

confidence: 99%

Development of a simple and effective silver staining protocol for detection of DNA fragments

Liu

Ayalew

et al. 2017

Electrophoresis

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“…In order to further detect the quality of the extracted longan DNA, SSR primers ( Figure 3) were used to amplify genomic DNA of longan; PCR conditions were as described by Benbouza et al (2006). PCR products were silver stained according to the steps described by Liang et al (2014). The results (Figures 2 and 3) show that extracted DNA was suitable for PCR analysis.…”

Section: Resultsmentioning

confidence: 97%

“…Preparation of polyacrylamide gel and electrophoresis of PCR products were carried out according to Benbouza et al (2006). Gels were silver stained according to the steps described by Liang et al (2014).…”

Section: Extraction Protocol For Genomic Dnamentioning

confidence: 99%

A valid measure to eliminate the influence of polysaccharides and polyphenols in recalcitrant longan (Dimocarpus longan L.) during DNA isolation

Liang¹,

De-qiang²,

Wen³

et al. 2015

Afr. J. Biotechnol.

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Large amounts of polysaccharides, polyphenols, tannins, proteins, and other secondary metabolites in recalcitrant longan leaves make it difficult to obtain high quality genomic DNA during extraction. To obtain good quality of nucleic acids from local longan leaves and for its downstream applications, a new protocol was developed. It consists of rapid isolation of stable nuclei, which hinders covalent interactions with phenolics, followed by DNA extraction. The yield and quality of the resulting DNA were satisfactory and suitable for PCR analysis and digestion with a restriction enzyme. Here, a valid combination measure (β-mercaptoethanol, PVP40 and PVPP were used at different stages) was created to eliminate the influence of polysaccharides and polyphenols in recalcitrant longan during DNA extraction, which will facilitate the development of molecular quantitative genetics of longan.

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“…DNA bands were visualized directly in a white light box, then photographed or scanned. The gel plate was agitated in a shaker throughout the staining process [17].…”

Section: Pcr-sscpmentioning

confidence: 99%

Allele mining in the caprine alpha-lactalbumin (LALBA) gene of native Saudi origin

El-Hanafy

Qureshi

Sabir

et al. 2016

Biotechnology & Biotechnological Equipment

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Alpha-lactalbumin (a-LA) is a distinctive whey protein found in bovine milk and the milk of other mammalian species. Its main function in the lactating mammary glands is involved in lactose biosynthesis, which makes it a potential quantitative trait locus. The aim of this study is to determine the incidence of single nucleotide polymorphisms (SNPs) within the exonic and intronic portions of the caprine a-LA gene (LALBA) in native Saudi breeds (Ardi, Habsi and Harri) in relation to utility traits. Blood samples were collected from 300 animals (100 for each breed). Genomic DNA was extracted and a 268 bp fragment of the LALBA gene comprising exon 3 and its flanking area was amplified. Subsequent digestion with MvaI restriction endonuclease resulted in two different banding patterns: aLA A1 /aLA A1 and aLA A1/aLA A2 , and two allelic forms, i.e. aLA A1 and aLA A2 . Nucleotide sequencing of the designated LALBA amplicons was done and, following successful BLAST analysis, the sequences were submitted to GenBank (NCBI Accession No. KM267632, KM267633, KM267634 and KP940442). SNPs were searched within breeds of Saudi origin and homology was sought among caprine, ovine and bovine species. A C > T transition was identified within the fifth nucleotide of the silent a-LA A2 allele. Phylogenetic analysis on the basis of LALBA nucleotide sequence of Saudi goats indicated similarity with evolutionarily related sheep, while the origin of bovine species and deer was located some distance away.

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A rapid and effective method for silver staining of PCR products separated in polyacrylamide gels

Cited by 20 publications

References 12 publications

Development of a simple and effective silver staining protocol for detection of DNA fragments

Development of a simple and effective silver staining protocol for detection of DNA fragments

A valid measure to eliminate the influence of polysaccharides and polyphenols in recalcitrant longan (Dimocarpus longan L.) during DNA isolation

Allele mining in the caprine alpha-lactalbumin (LALBA) gene of native Saudi origin

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