Description:
A key component of 2X SYBR PCR Reaction Mix is a
stringent hot-start DNA polymerase. This technology ensures
robust and highly sensitive PCR amplification and quantification.
It enables faster thermocycling and saves 53% running time over
standard protocol.
Application:
The mix contains Sybr® Green, which binds to double stranded DNA
specifically. It can be used in DNA quantification and two-step
Q-PCR. This reaction mix contains internal passive reference dye
(ROX). It is manufactured for the ABI 7500 PCR machine.
Recommended storage conditions:
This product should be stored at –20ºC. To avoid repeated
freeze-thaw, open vials should be kept at 4ºC.
Primer and probe design:
Appropriate software, such as ABI Primer Express™, should be
used.
1. Tm: 58~60°C with 17~25 nucleotides in length
2. To design a small amplicon, <150bp
3. To avoid secondary structures
4. To avoid more than 3 consecutive Gs
5. To avoid complementary 3’ ends
Recommended reaction conditions:
95°C, 5min -> (95°C, 5sec. -> 60°C, 30sec.) for 50 cycles ->
melting curve.
Notes:
Primer concentration should not be high. 100nM to 300nM
each
- primer usually gives best results
- Melting curve is useful way to distinguish nonspecific
amplification products and primer dimers from desired product.
It’s recommended to run melting curve following amplification
cycles.
- To do replicates
- To reduce pipetting error: preparing amplicon specific master
mix, using repeating pipet, and keeping pipetting volume in
manufacture suggested range are very effective to get tight Ct.
Reference:
Higuchi R, Dollinger G, Walsh PS, Griffith R; Bio/Technology 10:
413–417, 1992
This product is distributed
for laboratory research only.
Caution: Not for diagnostic use . |
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