高成功率PCR酵素『KOD FX』を用いた 糸状菌からのコロニーPCR
Contents
1. PCR DNA TKOD FX PCR 1 Aspergillus oryzae TE buffer 50 95C 10min 5 l 82 PCR PCR 1 5 PCR2. DNA SDS DNA RNase KODD FX PCR DNA PCH DNA www toyobo co jp bio 2 KOD FX KOD FX 200UX1 200 KFX 101 35 000 KOD FX 1U u oxPCR Buffer for KOD FX 200UX1 X 5 1 000 KFX 101X5 140 000 mM dNTPs 200Ux1 x10 2 000 1 KFX 101X10 60 000 50 KOD FX DNA TA
3. PCR grade water 6 ul x PCR buffer for KUD FX 5 ul mM dNTPs 10 Ll 10pmol ul Primer 1 1 5 ll 1 Aspergillus oryzae 10pmol ul Primer 2 1 5 ul ul KOD FX 1 0U ul 1 Ul Total reaction volume 50 Ll Target ITS 1 150bp 470bp Primer 1 GTAACAAGGT T C TCCGT Primer 2 CGTTCTTCATCGATG OAD TD PCR 84C min Seac 50 TE buffer 95C 10min O 55 Tmin 30 cvoles 0 ii 5 PCR Taq PCR PCR 30 Aspergillus O7yZg 2 PCR 1 0 kb 1 0 kb 0 5 kb 0 5 ko 0 1 kb 0 1 kb 3 A ogryzgg PCR 4 PCR M
4. TArget Clone Plus KOD TA TArget Clone Plus
5. P CR KOD FX PCR KOD FX PCR KDD DNA Polymerase PCR PCR DNA PCPH TKOD FX DNA PCHR PCRH Aspe g7 u5 oOyZg g 2005
6. 100bp ladder M 100bp ladder 6 Pichia pastris 1 KOD FX 1 Aspergillus oryzae 7 Penicillium camemberti 2 A PCR 2 Aspergillus niger 8 Penicillium requeforti 3 B Long PCR 3 Aspergillus terreus 9 Magnaporthe salvini 4 Saccharomyces cerevisiae 10 Schizophyllum commune 5 Schizosaccharomyces pombe 11 Sagenomella oligospora 12 Thysanophora penicillioides PCR 2 5 3 KOD FX 1 4 DINA KOD FX DNA PCR Taq PCP KOD FX
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