LiliF Diagnostic ProductsWe will share various news about iNtRON

RT/RT-PCR

ONE-STEP RT-PCR PreMix Kit

Cat.No Capacity Inquire
25101 50 rxn. Inquire
PRODUCT INFORMATION
Description

A master mix type RT-PCR PreMix Kit capable of performing cDNA synthesis and PCR reaction at once using gene specific primers

  • • One-step RT-PCR
  • • Ready to Use
       - All components required for RT-PCR reaction are in a single tube with solution type
       - Easy use by adding RNA, gene specific primers and DW only
  • • Sensitive cDNA synthesis and high specific PCR amplification
  • • Adjustable reaction volume according to requirement
  • • High specificity and accuracy by hot-start Taq DNA Polymerase

ONE-STEP RT-PCR PreMix Kit is a master mix type ONE-STEP RT-PCR PreMix Kit that can perform cDNA synthesis and PCR reaction all at once using gene specific primers. The general RT-PCR method is a two-step sequential reaction, cDNA synthesis using reverse transcriptase, oligo dT / random primers and PCR process with DNA polymerase. The method is suitable for gene expression studies, but it has drawbacks such as cross contamination in complex processes. Gene specific primer (GSP) is preferred for the confirmation of the existence of specific genes and quantitative / qualitative analysis of the expressed genes. It is important to minimize errors in the reaction by simplifying the experimental procedure. The ONE-STEP RT-PCR PreMix Kit was designed to synthesize first-strand cDNA from total RNA or mRNA template and do PCR in a tube. The product includes all components, reverse transcriptase and DNA polymerase, for each reaction with mixed solution type. The OptiScriptTM RT System of ONE-STEP RT-PCR PreMix Kit makes accurate cDNA and high specificity PCR from 1pg to 2mg of template RNA. The ONE-STEP RT-PCR PreMix Kit is optimized product to confirm the expression of low copy gene, and its stability is maintained for the first time under frozen condition for more than 1 year.

Applications
  • 01RNA qualification study
  • 02RNA expression study
  • 03Specific RNA detection
  • 04Virus research
  • 05Molecular diagnostic study
  • 06 Real-time RT-PCR, etc.
Kit Contents
Content 25101(50 rxn)
ONE-STEP RT-PCR PreMix Kit 200 ml x 2 tubes
Manual 1 ea

[ ONE-STEP RT-PCR PreMix Kit ]

Component in 20 ml reaction

  • • OptiScript™ RT System
  • • i-StarTaq™ DNA Polymerase
  • • RT-PCR Buffer(10x)
  • • dNTP Mixture
  • • Stabilizer
Technical Data

Observation of sensitivity and specificity through virus screening


 

After the decimal dilution of 108.0EID50 / 0.1ml of the uroepithelium, RNA was extracted with Viral Gene-spin™ Viral DNA / RNA Extraction Kit (Cat. No. 17151). Then first strand cDNA synthesis and PCR reaction were performed using ONE-STEP RT-PCR PreMix Kit. As can be seen from the result, it has high efficiency and specificity, so it can be used to detect various diseases.

Lane M, 100bp Ladder DNA Marker; lane 1, 10-2 희석; lane 2, 10-3 희석; lane 3, 10-4 희석; lane 4, 10-5 희석; lane 5, 10-6 희석; lane 6, 10-7 희석; lane 7, 10-8 희석; lane 8 : 10-9 희석

Observation of sensitivity and specificity


 

Total RNA was isolated from human cancer cell lines (SNU 5) using the easy-BLUE™ Total RNA Extraction Kit (Cat. No. 17061), and then synthesized each first strand of cDNA using AMV Reverse Transcriptase and Optiscript™ RT System. After the cDNA mixture was diluted to an appropriate concentration, the expression levels of β-Actin (400 bp) gene were compared. As can be seen from the above results, we found that Optiscript™ RT System is more sensitive than AMV RTase. Therefore, ONE-STEP RT-PCR PreMix Kit has high efficiency and specificity due to Optiscript™ RT System and i-StarTaq™ DNA Polymerase. In other words, OptiScript™ RT System allows sensitive reverse transcription reaction and i-StarTaq™ DNA Polymerase provides high specificity amplification as hot-start PCR enzyme.

TroubleShooting Guide
QMaxime products include loading dye. Does the product include loading dye too?
AONE-STEP RT-PCR PreMix is master mix type, and loading dye is not included.
QIn contents, RNase inhibitor is not included in. Do I have to purchase it separately?
AThis product does not contain RNase inhibitor. We are selling 2,000U RNase inhibitor.
QThere is no mention of the amount of template in product manual. How much should I use?
AYou can use template less than 500ng in case of total RNA, and 0.05-0.1ug for poly (A) RNA.
Related Product
관련제품 Product,No,Capacity,Inquire list
Product Cat.No Capacity inquire
Maxime™ RT-PCR PreMix 
25131 96 tubes Inquire
Maxime™ RT PreMix (Oligo (dT)15 Primer) 
25081 96 tubes Inquire
Power cDNA Synthesis Kit 
25011 30 rxn. Inquire
easy-BLUE™ Total RNA Extraction Kit
17061 100 ㎖ Inquire
17061L 100 ㎖ x 5 Inquire
easy-spin™ Total RNA Extraction Kit
17221 50 col. Inquire
RNA-spin™ Total RNA Extraction Kit
17211 50 col. Inquire

List

TOP