MethylCode™ 亚硫酸氢盐转化试剂盒
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Applied Biosystems™

MethylCode™ 亚硫酸氢盐转化试剂盒

MethylCode™ 亚硫酸氢盐转化试剂盒是一种可将 DNA 样品中的未甲基化胞嘧啶转化成尿嘧啶而甲基化胞嘧啶维持不变的快速且简便的方法。经修饰 DNA 适用于下游分析(包括限制性核酸内切酶酶切、测序和微阵列)的了解更多信息
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货号数量
MECOV5050 次反应
货号 MECOV50
价格(CNY)
-
数量:
50 次反应
MethylCode™ 亚硫酸氢盐转化试剂盒是一种可将 DNA 样品中的未甲基化胞嘧啶转化成尿嘧啶而甲基化胞嘧啶维持不变的快速且简便的方法。

经修饰 DNA 适用于下游分析(包括限制性核酸内切酶酶切、测序和微阵列)的 PCR 扩增。使用 MethylCode™ 亚硫酸氢盐转化试剂盒,您可以预期:

–简单、快速的胞嘧啶甲基化检测

–在大多数实验条件下的一致性
–得益于偶联 DNA 变性/亚硫酸盐处理步骤,未甲基化的胞嘧啶转化为尿嘧啶得到简化
–适用于后续基于分子的分析的超纯 DNA



试剂盒规格:

–方案:快速对 DNA 进行热变性,然后使用离心柱纯化法进行亚硫酸氢钠修饰

–样品来源:质粒、基因组和核酸内切酶酶切 DNA
– 输入 DNA:500 pg-2 μg DNA/处理,最佳为 200-500 ng
– 回收体积:在 10 μL 洗脱缓冲液中回收经纯化和亚硫酸氢盐处理的 DNA



内容物和储存:
MethylCode™ 亚硫酸氢盐转化试剂盒包含对 DNA 进行转化以进行甲基化研究所需的所有试剂。该试剂盒包括 CT 转化试剂、稀释缓冲液、重悬缓冲液、结合缓冲液、洗涤缓冲液、脱磺化缓冲液、洗脱缓冲液、离心柱和收集管。

可在室温下储存长达 24 月。
仅供科研使用。不可用于诊断程序。
规格
产品线MethylCode™
产品类型试剂盒,亚硫酸氢盐转化
数量50 次反应
样品类型DNA(基因组)
运输条件室温
原始材料细胞数量纯化基因组 DNA - 200 ng 至 500 ng
产品规格试剂盒
Unit SizeEach
内容与储存
MethylCode™ 亚硫酸氢盐转化试剂盒包含对 DNA 进行转化以进行甲基化研究所需的所有试剂。该试剂盒包括 CT 转化试剂、稀释缓冲液、重悬缓冲液、结合缓冲液、洗涤缓冲液、脱磺化缓冲液、洗脱缓冲液、离心柱和收集管。

常见问题解答 (FAQ)

I think my DNA may be under-converted (incomplete). What can you suggest?

Ensure that the DNA that you use with the MethylCode Bisulfite Conversion Kit is ultra clean. If particulate matter is present after adding the CT Conversion Reagent, then centrifuge the material at high speed and perform the conversion with the clear supernatant. Also, ensure that all of the liquid is at the bottom and not in the cap or walls of the PCR tube, prior to performing the conversion reaction

How can I quantify my DNA after bisulfite conversion?

Checking your converted sample by O.D.260/280 tends to be inconsistent. We suggest performing qPCR or agarose gel electrophoresis and comparing your sample to a known amount of DNA to quantify your DNA after bisulfite conversion.

How should I store my converted DNA after it is eluted from the column?

The converted DNA is stable for one day at room temperature, one week at 4 degrees C, and two to four months at -20 degrees C. We recommend on storing your converted DNA below -70 degrees C whenever possible (remember: the DNA is single-stranded and inherently less stable than dsDNA).

How long can I store the CT Conversion Reagent in the MethylCode Bisulfite Conversion Kit after it has been dissolved?

The CT Conversion Reagent that has been prepared in solution may be stored up to 1 week at -20 degrees C. We recommend thawing at room temperature and mixing for 2 mins by rotating or vortexing before use.

My PCR post-bisulfite conversion failed. What can you suggest?

Here are some recommendations:

- Primers: Ensure that your primers are designed to amplify the converted template. We recommend primers that are 24-32 nts in length and contain no more than 2-3 mixed bases (for base-pairing to C or T residues). The 3' end of your primer should not contain a mixed base or end in a residue whose conversion state is unknown.
- Polymerase: We recommend using a hot-start Taq polymerase such as Platinum Taq DNA Polymerase, Platinum Taq High Fidelity, or AccuPrime Taq DNA Polymerase. Proof-reading polymerases are not recommended because they cannot read through uracil present in DNA templates.
- Amplicon size: Bisulfite modification is harsh and may cause strand breaks. Most research publications recommend 200 bp lengths. Larger amplicons can be generated, but this requires an optimized protocol.
- Template DNA: We recommend using 2-4 µl of eluted DNA for each PCR reaction. Ensure that total template DNA is less than 500 ng.