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ADVANCEDtm cDNA SYNTHESIS KIT

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ADVANCED™ cDNA SYNTHESIS KIT
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介绍详情

 Ordering Information:

CAT.NO.

Quantity

Specification

801-100-XR

25 REACTIONS

cDNA synthesis

801-100-AR

100 REACTIONS

cDNA synthesis

Product Description: 

 Advantages:

Unbiased representation of 5’ and 3’ mRNA transcript ends

Sensitive detection of low copy number transcripts

High cDNA yields from as little as 1pg total RNA

Simple 2 tube system

Wisent cDNA Synthesis Kit uses the latest technology in reverse transcription buffer chemistry to enhance cDNA synthesis speed and yield with accurate transcript representation. The reverse transcriptase, buffer system and combination of random hexamers with anchored oligo(dT) allow for unbiased, efficient, sensitive cDNA synthesis.

High quality cDNA synthesis for downstream qPCR analysis is essential for successful expression studies. cDNA synthesis is affected by the reverse transcripase, buffer systems, enhancers and priming strategy. The Wisent cDNA synthesis mix removes the need for user optimisation of these critical factors.

Wisent (RTase) is both thermostable and extremely active. The enzyme is blended with RNase inhibitor.

The RTase is not inhibited by ribosomal and transfer RNAs, total RNA is an ideal substrate. The 5x cDNA synthesis mix can be used with up to 4.0μg total RNA. The relative concentrations of random hexamers and anchored oligo(dT) have been optimised for the generation of cDNA for use in real-time PCR experiments.

This mix is optimized to generate cDNA for downstream real-time PCR analysis.

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Product Stability And Storage:

On arrival store at -25°C to -15℃.

Avoid prolonged exposure to light.

If stored correctly the kit will retain full activity for 12 months. It is stable at 2-8°C for 1 month and can go through 30 freeze/thaw cycles with no loss of activity.

Instructions For Use:

OUR KIT CONTAINS :

5 x cDNA mix;anchored oligo(dT);random hexamers;

15 mM MgCl2, dNTPs, enhancers and stabilizers.

Do not add further enhancers or MgCl2 to the reaction.

Note:

Template: For total RNA: use between 4 pg and 4.0 ug per reaction.

Incubation temperature: For the majority of applications incubate with a temperature of 42 °C for 30 minutes . If regions of interest contain high secondary structure incubation temperatures of up to 55 °C

might be used.

Reaction Setup:

1 Before starting,allow 5 X cDNA mix to thaw and briefly vortex

2 Prepare a mastermix, as follows: - Important: insert reagents in sequence as listed

Add

Volume

Final concentration:

WISENT 5 x cDNA synthesis MIX 

4 ul

1X

20 x RTase

1.0 ul

- ADD BEFORE RNA

Total RNA

between 4 pg and 4 ug

 

Add PCR grade water up to 20 ul final volume

OPTIONAL no RT control set up: 

Add

Volume

Final  concentration

WISENT 5 x cDNA synthesis MIX 

4 ul

1X

Total RNA

between 4 pg and 4 ug

variable

Add PCR grade water up to 20 ul final volume

3 Program your instrument using following conditions, as follows

Cycles

Temperature

Time

Notes

1

42°C

30min

Incubation

1

85°C

10min

Enzyme denaturation

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