cDNA Amplification by SMART-PCR and Suppression Subtractive Hybridization (SSH)-PCR

Hillmann, Andrew; Dunne, Eimear; Kenny, Dermot

doi:10.1007/978-1-59745-553-4_15

Cited by 27 publications

(19 citation statements)

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“…Next, the cell was sucked into the recording pipette (supplementary material Fig. S1), and the cellular material was expelled into a tube for the SMART-PCR cDNA synthesis and amplification (Hillmann et al, 2009) (see Materials and Methods) followed by PCR with gene-specific primers to detect certain transcripts. In addition to CASR and GPRC6A, each attested cell was analyzed for the expression of NTPDase2, SNAP-25, PKD2L1, TRPM5, gustducin and T1R3.…”

Section: Single-cell Profiling Of Rna Transcriptsmentioning

confidence: 99%

Functional expression of the extracellular-Ca2+-sensing receptor in mouse taste cells

Bystrova

Romanov

Rogachevskaja

et al. 2010

Journal of Cell Science

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Section: Single-cell Profiling Of Rna Transcriptsmentioning

confidence: 99%

Functional expression of the extracellular-Ca2+-sensing receptor in mouse taste cells

Bystrova

Romanov

Rogachevskaja

et al. 2010

Journal of Cell Science

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“…which have been proven to play critical roles in the process of spermatogenesis and steroidogenesis. Various approaches have been used to analyze these tissueexclusively expressed genes including suppression subtractive hybridization (SSH), 8) differential display reverse transcription-polymerase chain reaction (RT-PCR), 9) microarray, 10) and whole genome approach analysis. 11) The UniGene database (http://www.ncbi.nlm.nih.gov/unigene) is a collection of sequences of individual clones from the cDNA libraries of various organs.…”

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confidence: 99%

Comparative and Functional Analysis of Testis-Specific Genes

Liu¹,

Jin²,

Li³

et al. 2011

Biological & Pharmaceutical Bulletin

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The testis is the special male gonad responsible for spermatogenesis and steroidogenesis with complex gene expressions. Characterizing and comparing the testis-specific genes in different species can reveal key genes related to testis specifical functions and provide supplementary information for study of human testis function. We screened testis-specific genes from Unigene libraries, total 317, 449 and 147 testis-specific genes were identified for human, mouse and rat, respectively. Ten from thirteen selected human testis-specific genes were validated exclusively expressed in the testis by reverse transcription polymerase chain reaction (RT-PCR). Systematic bioinformatics analysis showed that specific genes were mainly related to spermatogenesis and testis development process with significant Glycolysis and Pyruvate metabolism. Enrichment functions were discussed.Key words testis; specific; digital differential display Biol. Pharm. Bull. 34(1) 28-35 (2011) © 2011 Pharmaceutical Society of Japan * To whom correspondence should be addressed. e-mail: zxsys008@126.com Colon 7 29584 1 5219 3 5458 Heart 4 19926 8 67542 4 31600 Kidney 6 49568 10 54164 4 24601 Liver 11 72628 6 25033 3 59093 Lung 7 46559 7 40355 2 8065 Muscle 2 6424 2 3031 1 2429 Ovary 4 10862 2 6847 1 4443 Pancreas 1 4308 6 22020 1 16173 Placenta 12 71806 7 21182 2 8475 Prostate 5 63083 0 0 1 9076 Spleen 2 36689 3 46301 1 3629 Stomach 11 36759 4 18360 0 0 Thymus 4 72916 8 93976 0 0 Uterus 5 58541 0 0 0 0 Total 96 783531 74 454582 30 198862 Libraries from 18 organs against testis libraries were used in the digital differential display (DDD) program.

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“…Moreover, with only 2747 tumor-associated probe sets used in the study, our analysis reflected only a part of the altered gene expression patterns. In order to further understand the biological changes in 48A9 cells at the transcriptional level, we used an improved suppression subtractive hybridization (SSH) method to selectively determine differentially expressed genes[7]–[9]. Using this method in 48A9 cells exposed to spaceflight, we successfully identified several genes to be significantly up-regulated, including Src homology 3 domain-containing guanine nucleotide exchange factor ( SGEF ), submergence-1 ( SUB1 ), non-coding RNA metastasis-associated lung adenocarcinoma transcript 1 ( MALAT-1 ), cytochrome c oxidase subunit 2 ( MT-CO2 ), and myosin light chain 6 ( MYL6 ), and identified three new putative genes.…”

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confidence: 99%

Spaceflight alters the gene expression profile of cervical cancer cells

Zhang¹,

Tong²,

Wang³

et al. 2011

Chin J Cancer

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Our previous study revealed that spaceflight induced biological changes in human cervical carcinoma Caski cells. Here, we report that 48A9 cells, which were subcloned from Caski cells, experienced significant growth suppression and exhibited low tumorigenic ability after spaceflight. To further understand the potential mechanism at the transcriptional level, we compared gene expression between 48A9 cells and ground control Caski cells with suppression subtractive hybridization (SSH) and reverse Northern blotting methods, and analyzed the relative gene network and molecular functions with the Ingenuity Pathways Analysis (IPA) program. We found 5 genes, SUB1, SGEF, MALAT-1, MYL6, and MT-CO2, to be up-regulated and identified 3 new cDNAs, termed B4, B5, and C4, in 48A9 cells. In addition, we also identified the two most significant gene networks to indicate the function of these genes using the IPA program. To our knowledge, our results show for the first time that spaceflight can reduce the growth of tumor cells, and we also provide a new model for oncogenesis study.

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cDNA Amplification by SMART-PCR and Suppression Subtractive Hybridization (SSH)-PCR

Cited by 27 publications

References 4 publications

Functional expression of the extracellular-Ca2+-sensing receptor in mouse taste cells

Functional expression of the extracellular-Ca2+-sensing receptor in mouse taste cells

Comparative and Functional Analysis of Testis-Specific Genes

Spaceflight alters the gene expression profile of cervical cancer cells

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