PCR仪

文献中的引物：
Primers specific for unmethylated DNA were 5'-GTTATGTTATGTTTGTTGTATG-3' (sense, -1616 to -1594) and 5'-TAAAATCCACCAACACAATCA-3' (antisense, -1394 to -1414) and yielded a 222-bp PCR product.
Primers specific for methylated DNA were 5'-TACGTGTTAGGGTCGATCG-3' (sense, -1427 to -1409) and 5'-CGAAATATCTACGCTAAACG-3' (antisense, -1152 to -1171) and yielded a 276-bp PCR product.
1、不知文献引物是否可用。
2、设计甲基化引物是不是最好在5‘端的甲基化区（上面第一个甲基化区是5’端的，第二个是3‘端的）
3、还有一问题：
我用MethPrimer 设计，软件给出好几对甲基化引物，可不知该怎样选择，希望能赐教。能否帮我推荐一下cyclind2基因的甲基化引物。谢谢大家了。

请教:
我现在做MSP拉出的要么是引物二聚体,要么什么也没有,急急急啊,怎么办?请教各位大虾!

回答：1：改变PCR条件；
2：更换引物。
反复查阅文献，并不断实践。

刚开始做这个实验，请问您们用的是这个亚硫酸氢钠吗？要加水后反应生成亚硫酸氢钠？下面是说明：
S9000
Sigma-Aldrich Sodium bisulfite ReagentPlus®, ≥99%

As a solid, this product is sodium metabisulfite,.
When dissolved in water, it yields sodium
bisufite NaHSO3.
Na2S2O5 + H2O ® 2 NaHSO3
Sodium bisulfite is a reagent that is used in a variety of
large scale applications. These include the removal of
chlorine from bleached fibers in paper manufacture, as
a disinfectant and bleaching agent for wool, the
preparation of hot and cold indigo vats for dyeing, the
coagulation of rubber latex, and the manufacture of
sodium hydrosulfite.
请做过的朋友帮忙看一下，给点意见，谢谢！