SENO Taq DNA Polymerase PCR Kit 200T

運費美元 45 - 超過美元免費 300

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描述

規格

Taq DNA聚合酶 1000 U with mixed buffer (Shipping with icebag)

貯存 Conditions:

Taq DNA Polymerase and Taq 聚合酶鍊式反應 Kits should be stored at -30 to -15°C in a constant-temperature freezer upon receipt.

測試:

Taq polymerase

Notes Before Starting:

  • Provided with Load PCR Buffer containing gel-loading reagent and gel-tracking dyes.
  • PCR Buffer and Load PCR Buffer yield a final concentration of 1.5 mM MgCl2 in the reaction mix. Adjust Mg2+ concentration if necessary by adding 25 mM MgCl2.
  • High-quality PCR-grade dNTP Mix (10 毫米) is available separately if needed.
  • Keep PCR tubes on ice until placed in the thermal cycler.
  • Include a No Template Control (NTC) in every assay.

準備 and Mixing:

  • Thaw buffers and reagents at room temperature or on ice, then keep them on ice after complete thawing.
  • Prepare a reaction mix containing all PCR components except template DNA. Prepare 10% more volume than required.
  • Mix the reaction mix thoroughly and dispense it into PCR tubes.
  • Add template DNA (≤1 µg/reaction) to PCR tubes. For RT-PCR, add an aliquot from the reverse transcriptase reaction, not exceeding 10% of the final PCR volume.

Reaction setup using SENO Taq DNA Polymerase

成分Volume/reaction Final concentration
Reaction mix
10x PCR Bufferˢ¹ or 10 微升1X
10x Load PCR Buffer(選修的)
dNTP mix (10 mM of each)2 微升200 µM of each dNTP
Primer1Variable0.1–0.5 µM
Primer2Variable0.1–0.5 µM
Taq DNA聚合酶0.5 微升2.5 units/reaction
RNAse-free waterVariable
模板DNA (added at step 4)Variable≤1 µg/reaction
總反應體積 100 µlˢ²

Thermal Cycling:

Program thermal cycler according to manufacturer’s instructions. Refer to the provided cycling program.

Optimized cycling conditions for Taq DNA Polymerase

時間溫度Comment
Initial denaturation3 分分鐘94℃
3-step cycling:
變性0.5–1 min94℃
退火0.5–1 min50–68°CApproximately 5°C below Tm of primers.
擴大1 分分鐘72℃For PCR products longer than 1 知識庫, use an extension time of approximately 1 min per kb DNA.
Number of cycles25–35
Final extension10 分分鐘72℃

Simplified Hot Start:

Start PCR program. Once the thermal cycler reaches 94°C, place PCR tubes in the cycler for improved specificity.

Post-PCR:

  • After amplification, samples can be stored at 2–8°C overnight or at -20°C for longer storage.
  • PCR products can be directly loaded onto agarose gel using Load PCR Buffer without prior addition of loading buffer and gel-tracking dyes. Refer to the provided table for migration distance and gel tracking dyes.

Migration distance of gel tracking dyes in Load PCR Buffer

% TAE (TBE) Agarose 凝膠Red dyeOrange dye
0.8 500 (270) BP~80 (<10) BP
1.0 300 (220) BP〜40 (<10) BP
1.5 250 (120) BP 〜20 (<10) BP
2.0 100 (110) BP<10 (<10) BP
3.0 50 (100) BP<10 (<10) BP

附加資訊

重量0.7 公斤
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