Y-shaped linker, kit and method for constructing RRBS library
A kit and library technology, applied in the field of molecular breeding, can solve problems such as low enzyme digestion efficiency evaluation results, achieve the effects of improving its accuracy, avoiding mismatching, and improving accuracy
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Embodiment 1
[0044] (1) Sampling
[0045] According to the library building task list, carefully check whether the Novo number, sample name, and sample status of the sample are consistent with those marked on the library building list, place the samples in order, shake briefly after all the samples are melted, and centrifuge for sampling.
[0046] Prepare a 200 μL EP tube according to the library construction task, and add the corresponding volume of RNase-free H 2 O, according to the sampling volume of the library construction task list, take samples in sequence (3 μg genomic DNA for each sample), check the sample name and Novo number again when sampling, and take the sample after confirming that it is correct. After the sample is taken, the EP tube and the sample tube are one by one Correspondingly put it in the ice box, and continue to take the next sample. After all the samples are taken, check whether the sequence of the sample tubes is consistent with the sequence of the EP tubes. Af...
Embodiment 2
[0170] In this example, the same sample to be tested as in Example 1 was used, and the library was constructed according to the same steps, wherein the only difference was the Y-shaped joint, and the specific sequence information was as follows:
[0171] The first group: Compared with SEQ ID NO: 3, the methylated adapter 1 only lacks the base t complementary to the sequencing primer; compared with SEQ ID NO: 4, the methylated adapter 2 only lacks the base t complementary to the sequencing primer The same base a in the end base.
[0172] The second group: compared with SEQ ID NO: 3, the protected base sequence of methylated linker 1 is atg (reduced by half), compared with methylated linker 2 and SEQ ID NO: 4, the protected base sequence is tac (cut in half).
[0173] The third group: compared with SEQ ID NO: 3, methylated linker 1 has three tacs added at the end of the protected base sequence.
[0174] The fourth group: the methylated linker 1 is the same as SEQ ID NO: 3, and...
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