タイトルまたは説明
miRNA tailing dye method fluorescence quantitative PCR kit
Cat. No.: MR***2
Price: $**0/**0T (negotiable if bulk purchase)
Storage conditions: **0℃, valid for 1 year, avoid repeated
freezing and thawing
Product introduction
miRNA PolyA SYBR qPCR Kit uses LABLEAD's chemically modified
hot-start polymerase and SYBR Green I chimeric fluorescence method
for miRNA fluorescence amplification. It has high specificity and
can effectively avoid primer dimer formation and other forms of
non-specific amplification. The reaction buffer is fully optimized,
the PCR amplification efficiency is high, the fluorescence signal
is strong, and the reaction sensitivity is better than many similar
products. The kit is equipped with an optimized Tag Primer for
free, which makes the amplification effect of miRNA more
guaranteed.
Product features
The chemically modified hot-start polymerase is used, the reaction
specificity is high, and non-specific amplification can be
effectively avoided;
The reaction buffer is fully optimized, the PCR amplification
efficiency is high, and the reaction sensitivity is higher;
The optimized Tag Primer is provided free of charge, and the
amplification effect of miRNA is more guaranteed.
Product composition
|
component
|
**0T
|
**0T
|
**0T
|
|
miRNA SYBR Mastermix
|
1.0ml × 2
|
1.0ml × 3
|
1.0ml × 4
|
|
Tag Primer
|
*0ul
|
**0ul
|
**0ul
|
|
ddH2O
|
1.0ml × 2
|
1.0ml × 3
|
1.0ml × 4
|
|
U*-reward primer
|
*0ul × 2
|
*0ul × 3
|
*0ul × 4
|
Instructions:
1. Prepare the reaction system according to the table below:
Take an RNase-free centrifuge tube and prepare the following mixed
solution:
|
2× miRNA SYBR Mastermix
|
*0ul
|
|
Tag Primer（*0uM）
|
0.4ul
|
|
Specific primer（*0uM）
|
0.4ul
|
|
Temlate DNA/cDNA
|
0.2ul
|
|
ddH2O
|
to *0ul
|
* You can design and synthesize miRNA forward primers yourself.
Note: 1. Generally speaking, a final concentration of 0.2μM primers
in the reaction system can achieve a better amplification effect.
When the reaction results are not ideal, the primer concentration
can be adjusted within the range of 0.**1.0μM.
2. The sample volume is generally not more than 2.0ul. It is
recommended to optimize and explore for the first experiment. You
can take 2.0ul RT stock solution and 2, 4, 8, *6, *2, and *4 times
dilutions for qPCR.
3. If the PCR results are not ideal, consider using the *-stage
temperature setting for amplification. The amplification conditions
are: *5℃ 5min; *5℃ *0S; *0℃ *0S; *0℃ *0S (collecting
fluorescence signals).
4. Avoid strong light exposure during reagent storage and
operation.
5. When preparing the reaction system, avoid excessive shaking to
prevent bubbles.
Quality control
Using the synthesized U6 as a sample, using this kit, adding A,
reverse transcription, taking 1/*0 cDNA product for qPCR, the CT
value is less than *0.
