KR123-FastKing One Step RT-PCR Kit-171220

FastKing One Step RT-PCR Kit ——For fast and sensitive one-step RT-PCR

FastKing One Step RT-PCR Kit
Cat. no. 4992294

Storage
Store at -30~-15°Cfor up to one year.

Introduction
FastKing One Step RT-PCR Kit allows both reverse transcription and
gene amplification to take place in a single tube, which avoids cross
contamination between samples and improves the sensitivity of detection.
The 25×RT-PCR Enzyme Mix contains King reverse transcriptase, which is
a high efficient reverse transcriptase expressed by engineering bacteria;
a further-modified hot start Taq DNA polymerase, which provides high
efficiency and accuracy for the amplify reaction; and RNase inhibitor. With
a special modified hydrophobic motif, King RTase gets a significant affinity
for RNA and facilitates transcription through of RNA templates, especially
for templates with high GC content or complex secondary structures.
The 2×FastKing One Step RT-PCR MasterMix contains appropriate ion
concentration, dNTPs and PCR enhancer. It could stabilize both RTase and
polymerases and keep their efficiency within the whole reaction process.

Materials required but not supplied

1. RNA templates
2. gene-specific PCR primers

Protocol

1. Thoroughly thaw the template RNA, gene-specific PCR primers,
   2×FastKing One Step RT-PCR MasterMix and RNase-Free ddH2
   O,
   centrifuge briefly and place them on ice.
2. Prepare a reaction solution according to the following table on ice:

3. Set up thermal cycler conditions according the following table

4. Analyze the PCR products using agarose gel electrophoresis.

Important Notes

1. RNA template could be total RNA or mRNA. TRNzol or RNAprep pure kits can be
   used to purify high-quality total RNA.
2. RNase contaminations should be avoided in one-step RT-PCR. Some measures
   can be taken as below:
   1) Wear a disposable gloves and respirator to avoid the RNase contaminations
   from skin and saliva
   2) Operate the RNA related experiments in an RNase-free environment using
   RNase-free apparatus and consumable items.
   3) Consumable items related with RT-PCR should be incubated in 0.1% DEPC
   solution at 37°C for 12 hours and sterilized for 30 min before use.
3. 25×RT-PCR Enzyme Mix should be centrifuged briefly before use. Pipet slowly
   and store at -30~-15°C immediately after use.
4. Completely mix the 2×FastKing One Step RT-PCR MasterMix and centrifuge the
   reagent to the botiom of the tube before use.
5. Only use gene-specific primers and use specific primers according to specific
   requirements of experiments.

KR123-FastKing One Step RT-PCR Kit-171220

All trademarks or registered trademarks appearing on this website are the property of their respective owners.

This product is for scientific research use only. Do not use in medicine, clinical treatment, food or cosmetics.

Need more info ? Contact us anytime. We’re here: Go2biotech
E-mail: maggie@go2biotech.com / morgan@go2biotech.com
Telephone:+86 755 8399 5017