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名称:SYBR Green PCR预混试剂盒
品牌:ACTGene 型号:SYBR Green PCR预混试剂盒
产地:TW 新旧:0
价格:0 数量:2
是否促销:否
浏览次数:3762
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简介:SYBRGreen Real Time PCR Premix 2X (No ROX)
Product Description :
SYBRGreen Real Time PCR Premix 2X (No ROX) is an optimized ready-to-use mastermix containing unique reaction buffer, dNTP, Taq DNA polymerase and SYBR Green I. The user can easily use it to detect and quantify the expression of target genes via SYBR Green I Real Time PCR.
This product is for real-time PCR instruments from Roche, Bio-Rad, Ceheid, Corbett Research, Eppendorf. If you are using real-time PCR instruments form ABI, please purchase ROX reference dye separately.
Specification :100 x 50ul reactions (2 x 1.25ml)
Storage Conditions : -20℃
Procedure :
1. The concentration of input DNA sample could be 0.05fg/ul - 500pg/ul depending on the real-time PCR instrument. No need to purify samples if they are RT-PCR products.
2. Take out SYBRGreen Real Time PCR Premix 2X (No ROX) from -20oC freezer and thaw it. Keep the tube on ice.
3. Reaction setup: (Add each component in order)
Real Time Premix 2x 25.0ul
PCR Forward Primer (10uM) 1.0ul
PCR Reverse Primer (10uM) 1.0ul
DNA Template 4.0ul
H2O 19.0ul
Total 50.0ul
* Use 2ul H2O to replace DNA template for no template control
4. Real-time PCR program:
Temp.( oC) Time (sec.) Cycle
95 30 1
95
60 10
30 35- 45
5. Analyze amplification curve according to the manufacturers' instruction manual of Real Time PCR instruments.
Guidelines for Real-time PCR primer design :
(1) The PCR product size is ideally to be 60- 200bp
(2) Length of primer: 18-30bp; GC%: 40-60%; Tm: 60-68oC
(3) At the 3' end, it should be avoided to be T, or more than 3 sequential G or C. As for primer dimers, the ΔG should be > -5.0 kcal/mol or less than 3 sequential complementary bases.
(4) Check PCR efficiency of each primer pair by Agarose gel.