取扱説明書 - 東洋紡ライフサイエンス事業部
Contents
1. 0 1 Diethylpyrocarbonate DEPC 37 12 121 30 8 RT PCR Total RNA tRNA mRNA rRNA RNA MagExtractor RNA Code No NPK 201F Magical Trapper Code No MGS 101 MagExtractor System MFX Total RNA 10 9 1 mm s PCR Tm2. RNase 11 PCR RT Random Primer RT 65 C 5 5 RT poly A tail RNA RT Oligo dT 20 PCR 2 1 10 pmo PCR PCR RNA RNA
3. cDNA RNase H cDNA ReverTra Ace ReverTra Ace cDNA First strand cDNA RT PCR ReverTra Ace w ReverTra Ace First strand cDNA RT Positive Control DNA PCR High fidelity PCR long PCR RT PCR ReverTra Dash ReverTra Ace DNA KOD Dash RT PCR RT PCR RNA cDNA
4. G3PDH Housekeeping Gene RNA RNA Oligo dT 20 Random Primer p 9 cCDNA DNA lt RT PCR DNase RNA RT Primer F 559 mRNA 1 1237 lt 1110 intron Primer R size bases 1634 90 129 90 92 193 104 2 Control Primer F R Positive Control RNA RNA Human G3PDH Glyceraldehyde 3 Phosphate Dehydrogenase in vitro 3 22 mer poly A tail
5. 1 cDNA ReverTra Ace cDNA 14 kb cDNA 2 ReverTra Ace w PCR DNA RT PCR 1 ReverTra Dash cDNA ReverTra Ace PCR KOD Dash 105 RNA 1ug 2 RT PCR RT PCR 1 RNA First Strand cDNA cDNA Second Strand cDNA PCR RNA 5
6. 3 G3PDH Housekeeping Gene mRNA Tumor promoting Phorbol Esters RNA 4 hG3PDH RNA Poly A 22mers AAAAAAAAAAAAAAAAAAAAAA dd TTTTTTTTTTTTTTTTTTTT Primer F Primer R Oligo dT 20 0 45 kb 3 RNA 4 RT PCR 1 DNA PCR 2 UV PCR
7. 2 Oligo dT 20 poly A tail mRNA RNA rRNA tRNA 3 Random Primer poly A tail Random Primer PCR Random Primer 30 C 10 9 7 RNA 1 RNase RT PCR RNase RNase RNA RNase 2
8. 67 ul 10 x PCR Buffer 1041 10 pmol ul 141 10 pmol gl 141 KOD Dash 141 Total Volume 100 ul PCR 1 kb 98C 10 sec 60 C 2 sec 30 74 C 30sec 1 98 C 10 sec 70C 30 sec X30 2 Fast PCR PCR 1 kb 30 sec 1 min 1 kb KOD Dash Taq O 2 7 2 ReverTra Ace DNA 1 20 ul
9. 5 5 1 ReverTra Dash ReverTra Ace w RNase Free H2O 11 X ul 5 x RT Buffer 4 dNTP Mixture 10mM 2ul RNase Inhibitor 10U u I 141 Primer Random Primer 25 pmol u I Oligo dT 20 10 pmol u I 1 10 pmol ul RNA Total RNA 1 g mRNA 10 100ng 1 Positive Control RNA fO copies 1 u I ReverTra Ace 1ul Total Volume 20 ul 30 C 10 min 42 C 20 min 99C 5min l 4C 5min l Random Primer 30 10 Positive Control RNA Control Primer R cDNA 99 5 PCR 6 2 POR 1 ReverTra Dash 1 20
10. 2 d DNA DNase I Negative Control PCR La UID AS 10 1 Takagi M Nisioka M Kakihara H Kitabayashi M Inoue H Kawakami B Oka M and Imanaka T Appl Environ Microbiol 63 4504 4510 1997 12 11 Code No ReverTra Ace lt gt 10 000U x 1 TRT 101 KOD Dash lt PCR gt 250U x 1 LDP 101 250U x 1 x5 LDP 101X5 250U x 1 x 10 LDP 101X10 RNase Inhibitor recombinant 2500U x 1 SIN 201 Random Primer 9mer 2 5 nmol FSK 301 RNA MagExtractor RNA Total RNA gt 10
11. 67 ul 10 x PCR Buffer 10 ul 10 pmol Ah 141 10 pmol yl 141 DNA 2 5 U 1l Total Volume 100 uI PCR 1 kb 94C 30sec 60C 30sec 30 72 C 1min Mg Mqg 1 Taq 1 kb 1 min 1 kb 8 6 1 Control Primer R RNA
12. g Primer 3 Reverse Transcription RT Q E ReverTra Ace Reverse Transcriptase Synthesis of First Strand cDNA cDNA 3 5 DNA Polymerase KOD Dash Y Synthesis of Second Strand cDNA ReverTra Dash Primer DNA Polymerase KOD Dash i Amplification 1 RT PCR 2 3 20 100 1 ReverTra Ace 100 ul 2 5xRT Buffer 25 mM Mg 400 ul 3 RNase Inhibitor 10 U ul 100 ul 4 dNTP Mixture 10 mM 200 gl 5 RNase Free H20 1200 ul 6 Oligo dT 20 10 pmol u I 100 ul 7 Random Primer 25 pmol u I 100 8 Control Primer F 10 pmol AD 50 ul 9 Control Primer R 10 pmol u I 50 ul 10 Positive Control RNA 10 copies ul 50 gl 11 KOD Dash 2 5 Ul ul 100 ul 12 10xPCR Buffer 1000 ul ReverTra Dash Oligo dT 20 5 dT 2o 3 Random Primer 5 dN e 3 Control Primer F G3PDH 5 ACCACAGTCCATGCCATCAC 3 20 mer Control Primer R G3PDH 5 TCCACCACCCTGTTGCTGTA 3 20 mer 3 G3PDH Control Primer F R 0 45 kb PCR Human G3PDH Rat Mouse Swine
13. 0 NPK 201F Magical Trapper gt RT PCR RT PCR Quick Master Mix lt Tth DNA Pol RT PCR gt ReverTra Plus ReverTra Ace KOD Plus RT PCR gt PCR ReverTra Ace qPCR RT Kit Realtime PCR Master Mix PCR gt SYBR Green Realtime PCR Master Mix PCR SYBR Green HH FH SYBR Green Realtime PCR Master Mix Plus PCR SYBR Green HH FH THUNDERBIRD Probe qPCR Mix 7 PCR gt THUNDERBIRD SYBR qPCR Mix SYBR Green PCR gt 13 200 1ml x 5 1ml x 5 1ml x 5 1ml x 1 1 67mlx 3 1ml x 1 1 67ml x 3 MGS 101 PCR 311 PCR 501 FSQ 101 QPK 101 QPK 201 QPK 212 QPS 101T QPS 101 QPS 201T QPS 201 HPCR jm 530
14. 8230 2 8 TEL 06 6348 3786 FAX 06 6348 3833 E mail order_lifescience toyobo jp 141 8633 10 2 TEL 03 6422 4819 FAX 03 6422 4951 E mail order_lifescience toyobo jp TEL 06 6348 3888 FAX 06 6348 3833 9 00 12 00 13 00 17 00 E mail tech_osaka toyobo jp URL http www toyobo co jp bio
15. Ideas amp Chemistry First Strand cDNA Synthesis Kit ReverTra Ace a Code No FSK 101 RT PCR Kit ReverTra Dash Code No PCR 401 TOYOBO CO LTD Life Science Department OSAKA JAPAN A3280K RT PCR RNA RT PCR x V IR 1 Reverse Transcription RT PCR Polymerase Chain Reaction RT PCR RNA mRNA RNA cDNA MMLV Molony murine leukemia virus
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