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BL698A Fluorescent Quantitative PCR Kit (5X1mL)
The Biosharp BL698A Fluorescence Quantitative PCR Kit (5X1mL) is a specialized reagent for Real Time PCR using the SYBR Green I chimeric fluorescence
Product details

Biosharp BL698A Fluorescent Quantitative PCR Kit (5X1mL)

Manufacturer introduction:

Biosharp BL698A Fluorescent Quantitative PCR Kit (SYBR Green qPCR Mix)

Brand:biosharp

Item number:BL698A

Specifications:5X1mL

This product is made usingSYBR Green IChimeric fluorescence methodReal Time PCRSpecial reagents for hot startTaq DNAPolymerasedNTPSYBR Green IReady to use once the reagents are pre mixed2x Mix, can targetcDNAPerform rapid and highly specific quantitative detection. This product is chemically modifiedHot Start Taq DNAPolymerase, in50 ℃The following is inactive, only95 ℃Under certain conditions, the enzyme's activity can only be fully restored after heating, ZMinimize the production of non-specific amplification products and improve fluorescence quantificationPCRThe accuracy of the reaction. The unique reaction buffer of this kit can achieve good amplification results in a wide range, higher detection sensitivity, and stronger signal.

Product components:

product number

product name

specifications

1

2×SYBR Green qPCR Mix

5x1ml

2

50×ROX Reference Dye

1ml

storage condition

After receiving this product, please place it immediately-20℃Store in the dark, valid for one year28℃Storage under conditions6MonthAvoid repeated freeze-thaw cycles.

instructions

1、 PreparationReal-Time PCRReaction system:

1Collect all reagents2×SYBR Green qPCR MixROX Reference DyeTemplates, primers, andRNase-Free ddH2ODissolve and thoroughly mix at room temperatureAvoid generating bubbles.After brief centrifugation, placed on icePrepare the reaction solution according to the table below.

Refer to the table below to prepare the reaction system:

component

20ulsystem

final concentration

SYBR Green qPCR Mix

10μl

Primer F (10 μM)*

0.4~0.8 μl

0.2~0.4 uM*

Primer R (10 μM)*

0.4~0.8 μl

0.2~0.4 uM*

50 x ROX Reference Dye**

cDNAtemplate

--

RNase-Free ddH2O

to20 μl

* Generally speaking, the final concentration of primers in the reaction system is0.2 uMA better amplification effect can be obtained. When the reaction performance is poor, it can be achieved at the final concentration0.2~0.4 uMAdjust the primer concentration within the range.

** Z adaptability of several common instrumentsROX Reference DyeThe concentration is shown in the table below:

Instrument model

ppm

ABI PRISM 7000/7300/7700/7900HT/StepOne/StepOnePlusclass

ABI 7500/7500 FastQuantStudio ® 3/5QuantStudio 6/7 FlexViiA 7Stratagene Mx3000P/Mx3005PandMx4000class

RocheBio-RadEppendorfclass

No need to add

2Cover or seal the reaction tube/PCRPlate, gently mix well. Briefly centrifuge to ensure that all components are in the tube/Bottom of the board.

2、 Carry outReal-Time PCRreaction

1Suggest adopting a two-step approachPCRThe reaction program carries out the reaction.

Two-step reaction program:

step

temperature

duration

Number of cycles

Pre denaturation

95℃

5min

1

sex change

95℃

10sec

40

Annealing extension data acquisition

60℃

30sec

40

melting curve analysis

According to the recommended program settings for the instrument

equalFactors such as low template quantity can lead to poor amplification effect, and a three-step program can be used for this purposePCRReaction.

Three-step reaction program:

step

temperature

duration

Number of cycles

Pre denaturation

95℃

5min

1

sex change

95℃

10sec

40

annealing

50-60℃

30sec

40

Extended data collection

72℃

30sec

40

melting curve analysis

According to the recommended program settings for the instrument

2Machine detection: Place the reaction system in fluorescence quantificationPCRIn the instrument, run the program to collect data and analyze the results.

Biosharp BL698A Fluorescent Quantitative PCR Kit (5X1mL)

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