I startaq dna polymerase

Manufactured by iNtRON Biotechnology

I-StarTaq™ DNA Polymerase is a thermostable DNA polymerase enzyme used for polymerase chain reaction (PCR) amplification of DNA. It possesses 5'-3' DNA polymerase activity and exhibits high thermal stability.

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Lab products found in correlation

Fd0014, by Thermo Fisher Scientific (1 mentions) Fd0614, by Thermo Fisher Scientific (1 mentions) Safeview classic, by Applied Biological Materials (1 mentions) Licor 4300 sequencer, by LI COR (1 mentions) Rnaiso plus reagent, by Takara Bio (1 mentions) Maxime rt premix kit, by iNtRON Biotechnology (1 mentions) Geneamp pcr system 2700, by Thermo Fisher Scientific (1 mentions) Power sybr green pcr master mix, by Thermo Fisher Scientific (1 mentions) Quantitect reverse transcription kit, by Qiagen (1 mentions) Cfx96 real time pcr detection system, by Bio-Rad (1 mentions)

4 protocols using i startaq dna polymerase

Amplification and Genotyping of POU1F1 Gene

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The exon 6 and 3

^{'}

flanking region of POU1F1 gene was amplified
using F: 5

^{'}

-CCATCATCTCCCTTCTT-3

^{'}

and R: 5

^{'}

-AATGTACAATGTCCTTCTGAG-3)

^{'}

primers (Lan et al., 2007b). The 25 

µ L

PCR volume contained
100 ng genomic DNA, 0.5 

µ M

of each primers,

1 \times

 PCR
Buffer, 200 

µ M

dNTP, 2 mM

{MgCl}_{2}

and 1 U of Taq DNA
polymerase (i-StarTaq^™ DNA Polymerase, iNtRon Biotechnology). The
cycling protocol was 5 min at 95 

^{\circ}

C, 35 cycles of 94 

^{\circ}

C
for 30 s, 54 

^{\circ}

C annealing for 30 s, 72 

^{\circ}

C for 45 s with
a final extension at 72 

^{\circ}

C for 10 min.
PCR products of the POU1F1 gene were digested with 10 U of
AluI and PstI restriction enzymes (FD0014 and FD0614,
Thermo Fisher Scientific) at 37 

^{\circ}

C for 3 h. PCR products and
restriction fragments were electrophoresed on a 2.5 % agarose gel stained
with SafeView^™ Classic (Applied Biological
Materials Inc., Canada).
POU1F1 gene fragments which gave different genotypes were also
sequenced. The sequences of POU1F1 fragments were analyzed by using
the MEGA6 software (Molecular Evolutionary Genetics Analysis, version 6.0;
Tamura et al., 2013) for generating sequence alignments.

Işık R, & Bilgen G. (2019). Associations between genetic variants of the POU1F1 gene and production traits in Saanen goats. Archives Animal Breeding, 62(1), 249-255.

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Transposon Insertion-SCAR Genotyping

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The Mu-TD protocol was performed following methods described in Ramekar et al. (2018) (link). The specific primer sequences used are shown in Table 1. Amplification products were visualized using the gel system on a LI-COR 4300 sequencer according to the manufacturer’s protocol (LI-COR Biotech, Lincoln, USA). Fluorescently-labeled DNA (50–700 bp; 50–700 sizing standard LI-COR) served as molecular weight markers. Polymorphic parental lines were used to genotype the RIL population using 34 transposon insertion-SCARs previously developed by Roy et al. (2017) (link). The PCR mixture (20 μl) contained 20 ng of template DNA, 10× PCR buffer, 0.2 mM of each dNTP, 0.5 μm of the forward and reverse primers, and 0.025 U of i-Star Taq DNA polymerase (Intron Biotechnology, Korea). DNA was amplified using the following protocol: pre-denaturation at 95°C for 5 min; then 20 cycles of denaturation at 95°C for 30 s, then primer annealing at 56°C for 30 s, extension at 72°C for 1 min, and final extension at 72°C for 5 min to ensure complete extension of the PCR products. Amplicons were analyzed via gel electrophoresis with a 1.0% agarose gel. DNA fragments were visualized by ethidium bromide staining under UV light. Because both marker systems are dominant, they were manually scored as a binary response variable, with 1 (presence) and 0 (absence), respectively.

Ramekar R.V., Sa K.J., Park K.C., Roy N., Kim N.S, & Lee J.K. (2018). Construction of genetic linkage map and identification of QTLs related to agronomic traits in maize using DNA transposon-based markers. Breeding Science, 68(4), 465-473.

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Epididymal Fat RNA Isolation and qPCR Analysis

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Total RNA was isolated from the epididymal fat samples of the experimental mice using the RNAiso Plus reagent (Takara Bio Inc., Shiga, Japan) according to the manufacturer's instructions. cDNA was synthesized from 1 µg of the total RNA in a 20 µL reaction volume using a Maxime RT PreMix kit (iNtRON Biotechnology, Seongnam, Korea) containing the OptiScript™ reverse transcriptase and i-StarTaq™ DNA polymerase, following the manufacturer's recommended protocol. The oligonucleotide primers are shown in Table 2. The PCR conditions consisted of an initial denaturation step at 95°C for 5 min, followed by 30 amplification cycles consisting of denaturation for 40 s at 95°C, annealing for 40 s (temperature 56–62°C), and extension for 1 min at 72°C. The PCR products were separated on an agarose gel (1.5%) by electrophoresis for 30 min at 100 V. The bands were visualized, and their relative intensities were analyzed using the ImageJ software (National Institutes of Health, Bethesda, MD, USA).

Hong H., dela Cruz J.F., Kim W.S., Yoo K, & Hwang S.G. (2018). Glehnia littoralis Root Extract Inhibits Fat Accumulation in 3T3-L1 Cells and High-Fat Diet-Induced Obese Mice by Downregulating Adipogenic Gene Expression. Evidence-based Complementary and Alternative Medicine : eCAM, 2018, 1243049.

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Quantitative Analysis of Redrum Gene Expression

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Total RNA was isolated from indicated tissues of wild type and Redrum KO mouse. Reverse transcription was carried out with 1 μg of RNA from each sample using QuantiTect Reverse Transcription Kit (QIAGEN) according to manufacturer’s instructions. Standard PCR was performed on Gene-Amp PCR system2700 (Applied Biosystems) using i-StarTaq DNA polymerase (iNtRON Biotechnology). Real-time PCR for quantification was carried out using Power SYBR Green PCR Master Mix (Applied Biosystems) and CFX96^™ Real Time PCR Detection System (BIO-RAD). The following oligonucleotide primers were used for conventional PCR and real-time PCR: Redrum: (forward) 5′-AGACTGCTTTCTCAAGTGGCTTA G-3′, (reverse) 5′-AGCCAATGAATAGTCACTGTAGGG-3′; GAPDH: (forward) 5′-CAAGAAGGTGGTGAAGCAGG-3′, (reverse) 5′-CCGTATTCATTGTCATACCAGG-3′.

Lee Y., Park C., Lee S., Lee D, & Kim J. (2017). Isolation and Functional Examination of the Long Non-Coding RNA Redrum. Molecules and Cells, 41(2), 134-139.

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