Hot Start PCR

Paul, Natasha; Shum, Jonathan; Le, Tony

doi:10.1007/978-1-60761-629-0_19

Cited by 34 publications

(21 citation statements)

References 22 publications

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“…The smart technology, which is considered the most advanced approach to obtain the full length cDNA of a certain gene with a partial known sequence, was chosen for the full length cDNA of the HTA gene (11). At the same time, hot start PCR polymerase was applied to avoid non-specific amplification in lower temperatures (14) and a touch-down PCR program was applied to avoid non-specific amplification in early cycles and to improve the efficiency of specific amplification in later cycles of the program (15).…”

Section: Discussionmentioning

confidence: 99%

Molecular clone and functional study of a novel hepatoma associated gene

Liu

Zhao²,

Ju³

et al. 2013

International Journal of Oncology

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HTA is a novel hepatoma associated gene screened by bioinformatic strategies in our previous study. In the present investigation, the full-length sequence of the HTA gene was cloned by 3'-rapid amplification of cDNA ends (RACE) and 5'-RACE, which was 1414 bp and the open reading frame (ORF) was constituted with 3 exons and 2 introns. There are two types of splicing of the mRNA of HTA. Northern blot analysis showed that the 1.4 kb HTA mRNA and 1.7 kb HTA mRNA transcripts were present in the hepatocellular carcinoma (HCC) cell lines HepG2 and QGY-7703 but not in the normal cell line L02 and the human umbilical vein endothelial cells (HUVECs). Overexpression of the HTA gene in the hepatic cell line QSG-7701 via stable transfection can promote its proliferation rate and colony forming ability and change the cell cycle distribution of the cell lines. These results showed that the HTA gene is a potential therapeutic target in HCC and the clarification of its gene structure and sequence information provide an essential tool for future research.

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Section: Discussionmentioning

confidence: 99%

Molecular clone and functional study of a novel hepatoma associated gene

Liu

Zhao²,

Ju³

et al. 2013

International Journal of Oncology

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“…The amplification of partial 16S rRNA gene from two DNA samples was performed using hot start PCR technology (Paul et al, 2010) (JumpStart Taq Ready mix, Sigma Aldrich. Co. LLC, Bangalore, India) in order to avoid nonspecific amplification.…”

Section: Materials and Methods:-sample Collection:-mentioning

confidence: 99%

SPECIES IDENTIFICATION INFERRED THROUGH MITOCHONDRIAL 16S rRNA GENE SEQUENCE BETWEEN FRESHWATER TESTUDINES - LISSEMYS PUNCTATA AND MELANOCHELYS TRIJUGA.

Lalitha¹,

Chandavar.²

2017

IJAR

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“…However, these methods suffer from the unwanted characteristic that the amplification reaction is expected to proceed at room temperature if the nucleic acid sample is premixed with initiation reagents prior to compartmentalization. Conversely, in digital PCR the amplification reaction is controlled by temperature and any low-temperature non-specific pre-amplification process can be eliminated by using specific procedures [ 105 ]. As a consequence of that, the mixture can be compartmentalized prior to initiation with minimal risk of false positives if the infrastructure for an accurate temperature control is readily available.…”

Section: Isothermal Amplification Methodsmentioning

confidence: 99%

Isothermal Amplification Methods for the Detection of Nucleic Acids in Microfluidic Devices

2012

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Diagnostic tools for biomolecular detection need to fulfill specific requirements in terms of sensitivity, selectivity and high-throughput in order to widen their applicability and to minimize the cost of the assay. The nucleic acid amplification is a key step in DNA detection assays. It contributes to improving the assay sensitivity by enabling the detection of a limited number of target molecules. The use of microfluidic devices to miniaturize amplification protocols reduces the required sample volume and the analysis times and offers new possibilities for the process automation and integration in one single device. The vast majority of miniaturized systems for nucleic acid analysis exploit the polymerase chain reaction (PCR) amplification method, which requires repeated cycles of three or two temperature-dependent steps during the amplification of the nucleic acid target sequence. In contrast, low temperature isothermal amplification methods have no need for thermal cycling thus requiring simplified microfluidic device features. Here, the use of miniaturized analysis systems using isothermal amplification reactions for the nucleic acid amplification will be discussed.

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Hot Start PCR

Cited by 34 publications

References 22 publications

Molecular clone and functional study of a novel hepatoma associated gene

Molecular clone and functional study of a novel hepatoma associated gene

SPECIES IDENTIFICATION INFERRED THROUGH MITOCHONDRIAL 16S rRNA GENE SEQUENCE BETWEEN FRESHWATER TESTUDINES - LISSEMYS PUNCTATA AND MELANOCHELYS TRIJUGA.

Isothermal Amplification Methods for the Detection of Nucleic Acids in Microfluidic Devices

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