F305 R868 F18 R224 F91 R607 F369 R626 F50 R664 F306 R835 F1865 R2164 F1565 R1816 F56 R296
ctgggacgacataggagaaaa aaggaaggctggaagagtgc cacccgcgagtacaaccttc cccatacccaccatcacacc atatcgctgcgctggtcgtc aggatggcgtgagggagagc agaaggctggggctcatttg aggggccatccacagtcttc cagtgccagcctcgtctcat aggggccatccacagtcttc aggccggtgctgagtatgtc tgcctgcttcaccaccttct tgcggaaggatcattaacgga agtaggagaggagcgagcgacc cagccacccgagattgagca tagtagcgacgggcggtgtg aggggagagcgggtaagaga ggacaggactaggcggaaca
3
TIANGEN TRNzol-A+
RNA
5v/cm (20cm
rRNA
28SrRNA (
95v
1
)
RNA 80-85
28S(23S) 18S(16S)rRNA
18SrRNA
RNA
3)
3.2
RNA
1
700-1000ng RNA( 1-3ul) 5 V/cm 15-
20
rRNA RNA 80-85
28S(23S) 18S
RNA RNAsimple
RNA
2.2 / -
RNase TIANGEN
RNA RNAprep pure
RNA
237) RNA
TIANGEN
6
1
RNA
RNA
(
)
DNA
2
2.1
1
RNA
RNA
TIANGEN
RNA
TRNzol
TRNzol-A+
RNAplant
RNAsimple
ಢჟӬ৛࣮֒ࠥ!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!78
RNA
1 RNA
RNA
RNA RNA
RNA
RNA cDNA
PCR PCR
RT PCR
RNA Real-time PCR
1
2 RNA
2.1 RNA
2
2.2 RT-PCR/qPCR
28
‫ڼ‬ᅃቤ փཞᄣ೗RNA༵ൽํᄓ֡ፕ
28
‫ڼ‬ᅃবĂRNA༵ൽ‫ݛ‬ӄ
28
‫ܾڼ‬বĂRNA༵ൽ‫׉‬९࿚༶‫ݴ‬ဆ
38
‫ܾڼ‬ቤ RT-PCRํᄓ֡ፕ
39
‫ڼ‬ᅃবĂRT-PCRํᄓୁ‫ײ‬
39
‫ܾڼ‬বĂRTljPCR‫׉‬९࿚༶‫ݴ‬ဆ
43
‫ڼ‬ෙቤ ᆑ࠼ۨଉPCRํᄓ֡ፕ
44
‫ڼ‬ᅃবĂᆌᆩSYBR Green I݆৊ႜਨ‫ۨܔ‬ଉ‫ݴ‬ဆ
432bp 150bp 205bp 434bp 263bp 446bp
2
Human actin beta Rat actin beta Mouse actin beta human GAPDH Rat GAPDH Mouse GAPDH Rat 18s rRNA Human 18s rRNA Mouse 18s rRNA
RNase
2
RNaseH (RNaseH-)
RNA
cDNA
RNaseH
RNaseH
M-MLV
AMV RNA
DNA
16
RNA cDNA
RNaseH RNaseH
RNA RT
3
DMSO
RNA
20
20
10
45
0.4
10 DMSO
1/10 PCR
cDNA RNaseH
M-MLV RT-PCR
PCR
AMV
17
RT-PCR
1
-actin
RT-PCR
R actin f R actin r R actin f R actin r h actin f h actin r h actin f h actin r m actin f m actin r m actin f m actin r
tcaggtcatcactatcggcaat aaagaaagggtgtaaaacgca cccatctatgagggttacgc tttaatgtcacgcacgatttc tgacgtggacatccgcaaag ctggaaggtggacagcgagg cgggaaatcgtgcgtgac tggaaggtggacagcgagg gagaccttcaacaccccagc atgtcacgcacgatttccc gagaccttcaacaccccgc ccacaggattccatacccaa
1
/ RNA
2
/
RNA /
3 DNA/RNase
/ DNA
DNA
RNase /
DNA /
4
RNase RNA RNA
RNA
RNA RNA
RNase
DNase I(RT411)
DNA
RNA
8
RNA
RNA
RNA
Northern blot
RT-PCR
RNA
RNA
RNA RNA
cDNA
RNA
1 RNA
RNA RNA RNA
RNA
RNAprep pure RNAplant
TRNzol-A+ TRNzol RNAsimple RNase
RNase
2.3
RNA RNase RNA
RNA
2.4
RNA
RNA
RNA
TIANGEN RNAsafe
RNAsafe(DP409)
RNase
RNA
Tris MOPS
RNAsafe
RNase
ణ!!୤!
ጺஃ
1
1ĂٗRNA༵ൽ‫ڟ‬ᆑ࠼ۨଉঢ়‫ײୁۆ‬
1
2ĂٗRNA༵ൽ‫ڟ‬ᆑ࠼ۨଉׂ೗჋ስኸళ
2
3Ă“ٗRNA༵ൽ‫ڟ‬qPCRྜኝ঴ਦ‫ݛ‬ӄ”ׂ೗೅ದೠॏ
4
‫ڼ‬ᅃೊ RNA༵ൽĂRT-PCRĂᆑ࠼ۨଉ૙ஃև‫ݴ‬
5
‫ڼ‬ᅃቤ RNA༵ൽᇱ૙ᇑᆌᆩ
5
‫ڼ‬ᅃবĂRNA༵ൽୁ‫ײ‬
5
‫ܾڼ‬বĂRNA‫ࣅك‬तइ‫ڥ‬
44
‫ܾڼ‬বĂᆌᆩSYBR Green I݆৊ႜJunएᅺ‫ڦ‬՗‫ֶٳ‬ᅴ‫ݴ‬ဆ
47
‫ڼ‬ෙবĂᆌᆩTaqMan݆৊ႜਨ‫ۨܔ‬ଉ‫ݴ‬ဆ
50
‫຺ڼ‬বĂReal-time PCR‫׉‬९࿚༶‫ݴ‬ဆ
52
޹୤ᅃǖׂ೗ଚ՗!
65
޹୤ܾǖႎׂ೗ླྀ४!
67
޹୤ෙǖׂ೗၍߁ຎ!
72
‫ޜ‬ခኸళ!
76
TIANGENᆑ࠼ۨଉಢჟӬ!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! !!!!!!!77
(16S)rRNA 28S rRNA
18S rRNA
RNA
( 4) RNA
( 0.5 TBE)
0.5M NaOH 20
RNase
(kb) M 1 2 3
6 5 4
2.5 2 1.5 1
0.5
4
TIANGEN
RNA(
1
RNA
2
RNA
3
RNA
TRNzol 0.8%)
rRNA
18S 28S 18S 28S 18S 28S 16S 18S 23S 25S 18S 26S 16S 23S 18S 28S
16
‫ڼ‬࿵বĂRT-PCR‫׉‬ᆩా֖ᆅ࿿
18
‫ڼ‬ෙቤ ᆑ࠼ۨଉPCRरຍᇱ૙तຕ਍‫ݴ‬ဆ
19
‫ڼ‬ᅃবĂᆑ࠼ۨଉPCRຕბᇱ૙ࢅएԨ߁౒
20
‫ܾڼ‬বĂᆑ࠼ۨଉPCR‫ࣅڦ‬ბᇱ૙
22
‫ڼ‬ෙবĂᆑ࠼ۨଉᆅ࿿त༑ኍยऺᇱሶ
25
‫຺ڼ‬বĂᆑ࠼ۨଉPCR‫ڦ‬ᆌᆩ
27
‫ܾڼ‬ೊ RNA༵ൽĂRT-PCRĂᆑ࠼ۨଉํ७և‫ݴ‬
RNA RNAstore
RNA
(-20 -70
RNA
2 2.1
5
/
RNA
RNA
-
pH
RNA
RNA DNA
RNA
RNA
TRNzol-A+(DP421) TRNzol(DP405) RNAsimple(DP419)
/
RNA
TRNzol-A+ TRNzol
RNAsimple
RNA
TRNzol-A+ TRNzol
7
‫ڼ‬ෙবĂᅃၵ༬๺ፇኯ‫ڦ‬RNA༵ൽ
8
‫຺ڼ‬বĂRNAೠॏᇑ६ۨ
9
‫ڼ‬࿵বĂRNA‫ڦ‬ԍࢺ
11
‫ܾڼ‬ቤ RT-PCRᇱ૙ᇑᆌᆩ
13
‫ڼ‬ᅃবĂRT-PCR‫ݒ‬ᆌ༹ဣ
13
‫ܾڼ‬বĂRT-PCR‫݆ݛ‬
15
‫ڼ‬ෙবĂ༵ߛRT-PCR‫ݒ‬ᆌ‫ڦ‬ଳ௺‫܈‬ᇑ༬ᅴႠ
16
‫຺ڼ‬বĂRT-PCR‫ࠀׯ‬ᄲ໎
DNA
RNA
PCR
RNase
RNA
RNAstore TRNzol RNAprep pure
DMSO SDS