GF1060
GF1060-Fig1

2x SYBR Green qPCR Master Mix #GF1060

$160.00$380.00

Catalog#  GF1060

Quantity   4 ml of 2x master mix, enough for 400 20 µl-reactions, or 320 25µl-reactions

For research use only.

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SKU: GF1060. Category: .

Product Description

The 2x Gene-Foci® PCR master mix is a pre-mixed 2x solution of Taq DNA polymerase, PCR reaction buffer, dNTPs, MgCl2, enzyme stabilizer, PCR enhancers and SYBR® Green fluorescein. The master mix contains all the necessary components for qPCR amplification except template DNA and primers. This PCR master mix has been optimized for high-efficiency qPCR amplification, and is ideal for quantifying cDNA and genomic DNA.

Features:    

Super active and thermal-stable DNA polymerase in the master mix guarantees stable and robust amplification of target sequence.

PCR enhancers and enzyme stabilizers in the master mix make the qPCR reaction more reproducible.

Ultra sensitive, can detect trace amount of genomic DNA or cDNA.

Affordable, significantly cheaper than similar products from competitors.

Storage :     

For long term storage, keep at -80°C or -20°C. Shield against light. For short term storage, thaw the master mix and store at 4°C. The master mix is stable at 4°C for at least one month. Avoid repeated freeze and thaws.

 

 

 

Fig. 1. The cDNA synthesized using Gene-Foci® High Efficiency Reverse transcription kit was diluted 1:200, 1:500, 1:2000 and 1:5000, then 18S rRNA expression level was examined with Gene-Foci® SYBR® Green qCR Master Mix.

 

 

 

 

 

 

qPCR Reaction Setup

To setup a 25µl qPCR reaction, add the following components into a DNase-Free qPCR plate or tube:

Components

Volume

Final Concentration

2X Fast PCR Master Mix

12.5 µl

1X

Forward Primer (1 μM)

1~7.5 µl

50~300 nM[1]

Reverse Primer (1 μM)

1~7.5 µl

50~300 nM[1]

DNA Template

1~5 µl

0.1~10 ng/μl

ddH2O

to 25 µl

 

 

 

 

 

 

 

[1]   A primer titration from 50 nM to 300 nM final concentration should be performed to determine the     most amplification-efficient concentration.

qPCR Thermal Cycling Program

Set up the program qPCR on a qPCR machine as follows:

Option I: Two-step PCR

Cycles

Temperature

Time

(1) Pre-denature

1

95°C

3 minutes

(2) Thermal cycling

40

95°C

15 seconds

55~60°C(a)

30~60 seconds

 

Option II: Three-step PCR

Cycles

Temperature

Time

(1) Pre-denature

1

95°C

3 minutes

(2) Thermal cycling

40

95°C

15 seconds

55~60°C(a)

20 seconds

72°C

30 seconds

(a) Temperature required for annealing/extension step may need adjustment for different DNA templates and primers.