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RT-PCR Kit

FastKing RT Kit (With gDNase)

Efficiently read all kinds of sequences and accurately identify low abundance templates

Catalog number / packaging

Cat. no

ID

No. of preps

4992250

 KR116-03 1000 rxn

4992224

 KR116-02 100 rxn

4992223

 KR116-01 25 rxn
Manual Inquire Inspection report

  • Packing Size

    20 μl×25 rxn

  • Storage

    Store at -20℃.

  • Description

    The kit is an efficient, stable and fast reverse transcription system capable of removing genomic DNA contamination. The kit contains gDNase for efficiently removing genomic DNA, thus effectively avoiding interference of genomic DNA in total RNA. The high-efficiency reverse transcriptase FastKing RT Enzyme is a new type of modified reverse transcriptase, with a hydrophobic motif, which has stronger RNA affinity and thermal stability, thus further improving its reverse transcription efficiency and reaction rate, and making it more prominent in reading through RNA templates with high GC content and complex secondary structure and in stress resistance.

  • Features

    ■ High efficiency: The FastKing RT Enzyme is modified with hydrophobic motif, with RT efficiency more than 95%.

    ■ Sensitive: As low as 1 ng templates can be accurately identified.

    ■ Resistance: Capable of reverse transcription of complex templates, with perfect resistance to impurities.

    ■ Fast procedure: gDNA-free cDNA can be synthesized efficiently within 21 min.

  • Applications

    The reverse transcribed cDNA can be used in conventional PCR, Real time PCR, cDNA library construction.

Experimental Example

  • Group 1: Reverse transcription without gDNase treatment; Group 2: No gDNase treatment and no reverse transcription; Group 3: Reverse transcription after gDNase treatment; Group 4: gDNase treatment without reverse transcription. Methods: Fluorescence quantitative PCR detection of the TNF-alpha gene (primer designed on exon with cDNA or genome as template) using 1 μg Hela cell RNA (with genome residue) as template.

    Results: As shown in the figure, group 2 can reflect the residue of genome in RNA, group 3 can accurately reflect the true expression level of TNF-alpha, group 1 has errors in final quantitative results due to genome residue, and group 4 show that FastKing RT Kit can completely remove the residual genomic DNA in RNA.


  • 21 min reaction in one-tube

    It only takes 21 min to complete the gDNA removal and efficient reverse transcription process in the same tube without replacing the reaction tube and independent DNase I treatment process. Compared with the traditional method that requires 12-step operation and 140 min reaction, it greatly simplifies the operation steps and saves a lot of operation time.

  • Outstanding quality of King RTase

    Ultra-high reverse transcription efficiency

    ——Reverse transcription efficiency is over 95%

    The general reverse transcriptase has a reverse transcription efficiency of 40-60%, and cDNA yield can be increased by a higher RNA loading amount. King reverse transcriptase can achieve a reverse transcription efficiency of more than 95% due to its unique high affinity for RNA templates. Therefore, subsequent experiments can be satisfied without the need of a large amount of RNA input, which saves RNA and enables high purity and high yield of cDNA.




  • Easily read through complex templates

    ——Easily read through high GC and complex templates

    Single-stranded RNA has a wide range of complex secondary structure regions due to hydrogen bonding between strands. Ordinary reverse transcriptase may lead to termination of reverse transcription when encountering complex secondary structure, thus unable to successfully complete cDNA synthesis. However, the new generation of King reverse transcriptase has a unique structural domain, which can destroy the hydrogen bond between RNA strands, thus opening the complex secondary structure of RNA and ensuring the smooth reverse transcription.



  • Figure 1. Reverse transcription of mouse RNA was performed using TIANGEN FastKing RT Kit (left) and relevant product of Supplier A (right), then MM5 gene was quantitively amplified using TIANGEN SuperReal PreMix Plus (SYBR Green). The amplification curve and melting curve were analyzed. The RNA input was 1000 ng, 100 ng, 10 ng and 1 ng respectively. The results show that TIANGEN FastKing RT Kit has clear reverse transcription gradient and low Ct value, and has obvious advantages for reverse transcription of low abundance template (1 ng, blue arrow).


  • Figure 2. Reverse transcription of normal RNA template (red), template with large phenol residue (green) and template with alcohol residue (blue) of rats using TIANGEN FastKing RT Kit and relevant product of Supplier A respectively, quantify RNC genes using TIANGEN SuperReal PreMix Plus (SYBR Green), and amplification curves and Ct values were analyzed. The results show that TIANGEN FastKing RT Kit has the lowest quantitative Ct value after reverse transcription and excellent stress resistance, and has obvious advantages for templates with high impurity residues


Publication

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