DNA PCR Assays for Igh Rearrangement (original) (raw)

Bio-protocol, 2014

Abstract

1. Mouse spleen cells or ES-derived hematopoietic cells 2. DNA extraction Kit: PerfectPure DNA Cultured Cell Kit (5 PRIME, catalog number: 2302000) 3. 10x PCR Buffer with KCl (Life Technologies, Applied biosystems®) 4. MgCl2 (Life Technologies, Applied biosystems®) 5. Taq DNA polymerase (Life Technologies, Applied Biosystems®, catalog number: 4338856) 6. dNTPs (Life Technologies, Applied Biosystems®) 7. Primers (FASMAC) The sequence of primers are as follows. a. DHL(5’), GGAATTCG(AorC)TTTTTGT(CorG)AAGGGATCTACTACTGTG b. Mu0(5’), CCGCATGCCAAGGCTAGCCTGAAAGATTACC c. J3(3’), GTCTAGATTCTCACAAGAGTCCGATAGACCCTGG 8. Agarose (UltraPureTM Agarose) (Life Technologies, InvitrogenTM, catalog number: 16500-100) 9. Ethidium bromide (Wako Pure Chemical Industries, catalog number: 315-90051)

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