Method for extensively screening scallop SNP
A large-scale, screening technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problem of low separation efficiency of whole genome sequencing, and achieve the effect of excellent effect, safe and reliable principle, and reliable operation.
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[0030] The specific operation steps of this embodiment are as follows:
[0031] 1. Use guanidinium isothiocyanate, phenol, and chloroform to extract individual RNA from the blastula, gastrula, trochophore, D-shaped larva, and adult shellfish tissues of Chlamys farreri, and mix them in proportion to measure them by UV spectrophotometry. meter to measure its concentration for later use;
[0032] 2. Construction of full-length cDNA samples of Chlamys farreri:
[0033] Refer to the SMARTTM cDNA Library Construction Kit, but the sequence of the cDNA synthesis primer needs to be replaced with 5'-AAGCAGTGGTATCAACGCAGAGTCGCAGTCGGTACTTTTTCTTTTTTV-3'.
[0034] (1) cDNA first-strand synthesis:
[0035] For the reverse transcription reaction, use 1ug RNA as a template, add RNase-Free water to 4ul, add 1ul 10uM cDNA synthesis primers, 1ul 10uM SMART IVTM Oligonucleotide (5'-AAGCAGTGGTATCAACGCAGAGTACGCGGG-3'), incubate at 70°C for 3min, and place on ice immediately Cool for 1 min; then a...
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