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商品編號


P7640L



品牌


Enzymatics



公司


Enzymatics,inc



公司分類(lèi)


Kits



Size

1000 Reactions

商品信息

ZipScript? One-Step RT-qPCR Kit


Product Description

The ZipScript One-Step RT-qPCR Kit is a highly sensitive and reproduc
IBL
e RT-qPCR solution optimized for real-time PCR. The 25X enzyme mix is accompanied by a 2X reaction buffer. If a passive reference dye, such as ROX, is required for your assay, it can be purchased separately (P/N: Y9390).



Quality Control Analysis

The functionality of the ZipScript is evaluated by amplification of two mRNA transcripts in a one-step quantitative?RT-qPCR assay. The amplification threshold (Ct) of the test lot is compared to a reference lot.

Kit Contents:
25X ZipScript Enzyme Mix 2X ZipScript Reaction Buffer I

Notes:
Enzyme components were tested prior to formulation of the master mix and found free of contaminating endonucleases and exonucleases. Enzyme purity was >99% as determined by SDS-PAGE and neglig
IBL
e E.coli genomic DNA contamination was confirmed by qPCR. Specific activity was verified for each enzyme pre-formulation.

Protocol

ZipScript One-Step?RT-qPCR Reaction Setup:


All components and reaction set up should be kept on ice.

Thaw the 2X ZipScript Reaction Buffer I completely and vortex for 3-5 seconds to mix thoroughly. Quick spin to collect contents if necessary.

Prepare primer/probe mix. A final concentration of 0.4-0.9 ?M for each primer and 0.1-0.5 ?M for probe is recommended. However, the optimal concentration for primers/probe needs to be empirically determined for each assay.

Determine the number of reactions to prepare, including No Template Controls (NTCs). Add 10% extra volume to compensate for the pipetting loss.

Follow the table below to set up the reaction mix. It is recommended to make a master mix to minimize variations and potential errors.




Components

Volume/Rxn

Final Conc



Nuclease-Free Water

To a final 20 ?L reaction volume

–



2X ZipScript Reaction Buffer I

10 ?L

11X



50X ROX (optional)

0.4 ?L

1X



Primer/Probe Mix

X ?L

Variable



25X ZipScript Enzyme Mix

0.8 ?L

0.8 ?L



RNA Template

Variable

–





Seal PCR plate and spin briefly to bring down reagents.


Thermal Cycling Conditions:

Program the cycling conditions based on the recommendations below.

Standard Cycling Program*




Steps

Temperature

Time

Cycles



Reverse Transcription

50°C

15 min

1



Taq Activation/InitialDenaturation

95°C

2 min

1



Denaturation

95°C

15 min

40



Annealing/Extension*

60°C

30-60 sec





* Cycling parameters can be modified (especially the annealing/extension condition) to fit specific primer/probe selection.


View PDF Poster Instructions FAQ


Product Information



ZipScript? One-Step RT-qPCR Kit


Part Number
P7640L
Price
$1,338
Concentration
N/A
Unit Size
1000 Reactions


SDS

Available on request








Product Specification*


Storage Temperature
-25°C to -15°C






Assay

RT-qPCR


Specification
Amplification of Test Lot within 1Ct of Reference Lot for two different mRNA targets



* For a detailed summary of assay conditions and data, refer to the Quality Controls Analysis section.

Additional Downloads

Product Note



Limitations of Use
This product was developed, manufactured, and sold for in vitro use only. The product is not suitable for admi
NIST
ration to humans or animals. SDS sheets relevant to this product are available upon request.