RAPID Lite

STAT3

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I. DNA amplification

Kit
GE:illustra GenomiPhi V2 DNA Amplification Kit

II. PCR

Primer details

Exon No.	Forward sequence	Reverse sequence	Expected PCR product size (bp)	Genomic regions amplified by ePCR with the primer
2	CCTACTGGGACTTGTGGTGAAC	TCATGTCTCTTGACTCAAACTGAAAC	256bp	chr17:40500346-40500601
3	GCAAGAAGAATCTAACATGATGAATG	TCAGGTAATGATGCTCACGG	479bp	chr17:40498399-40498877
4	GGTGCCACAGTTCAGTCCAC	ACCATTGGGTCTGTTGGATTC	405bp	chr17:40497341-40497745
5	GGTAGTTAGGGTGGGATGGAAG	CCTGAGTGGCAGAGCAAGAC	270bp	chr17:40491246-40491515
6	AACAGGGAGCCTTCTCTTGG	ATGACCAGGCTCCTTTGAGG	382bp	chr17:40490606-40490987
7	GAGTCAAGGAGGCAAGTGAATATTAG	AACCAAGCAAGTTCTGCCAC	1063bp	chr17:40489697-40490759
8	GAAGTTCCTGCTCTGGAGTTG	AGTACCAATTCTGTGGGCCTG	324bp	chr17:40489368-40489691
9	GTGGTCTCCATGTCTTCAGTATTTC	CCCGCCTCTCACTCTACCAC	327bp	chr17:40485835-40486161
10	TCACGTGGTAGAGTGAGAGGC	CACCTGGAAAGAATGACCCTG	264bp	chr17:40485595-40485858
11	GACAGCTTGGCCTATTTACCTG	TGACAATGCACCCCAAGGCT	327bp	chr17:40483340-40483666
12-13	AGTAAATAACAGGTGGTCAAAGTAGG	AAAGGCAGGTGTCCTGTGAG	385bp	chr17:40481490-40481874
14	CACTTGGTATGTGGGAGGAGC	GAACAGGTTGATGTTTCTAATTCTGG	252bp	chr17:40481326-40481577
15	TGCTGCTTAGACTGGTCTCG	CAGTAATCATTCCACCTTCTCTTG	309bp	chr17:40478034-40478342
16	ATGTCCTTTCATTCTGAGCCC	TGTTTAGATGAGGGATGGTGC	292bp	chr17:40476888-40477179
17	CCTAGTATAGACAATGAGCACCATCC	TCAGTAGACATGGCCCAAATG	286bp	chr17:40476640-40476925
18	CCATTGTGGGATGAATGTGTG	ACCAAGAGTTCAGGGCCTTC	242bp	chr17:40475498-40475739
19	GTAGACGGGCCTGAGGATTTG	ATTGCTAACAGGGCATCCATC	279bp	chr17:40475192-40475470
20	TGCACAGATGTAACCAAGTCC	TGCTTGCAACTAGAAGCAGTG	311bp	chr17:40474950-40475260
21	GAACAGGGTGTTCAGGGTCTC	CCCAGGTTATTCAGGCATTTG	447bp	chr17:40474201-40474647
22	GCTTCCTCCAGCTCTGCTTAC	GTCCCATTCCCAGGGATAAC	179bp	chr17:40469133-40469311
23	TTCCATTGTGTCTTGTCAACC	CCAACTTCCCTTATAGGGACAAAG	215bp	chr17:40468763-40468977
24a	TGGATAGCAGTGGTAGCCTGG	GATCACCCACATTCACTCATTTC	360bp	chr17:40467566-40467925

III. Sequencing reaction

Kit
BigDye® Terminator v3.1 Cycle Sequencing Kit
Primer details

Exon No.	Forward sequence	Reverse sequence	Expected PCR product size (bp)	Genomic regions amplified by ePCR with the primer
2	CCTACTGGGACTTGTGGTGAAC	TCATGTCTCTTGACTCAAACTGAAAC	256bp	chr17:40500346-40500601
3	GCAAGAAGAATCTAACATGATGAATG	TCAGGTAATGATGCTCACGG	479bp	chr17:40498399-40498877
4	GGTGCCACAGTTCAGTCCAC	GATTTTCCATTCCTCCCAGA	218bp	chr17:40497528-40497745
5	GGTAGTTAGGGTGGGATGGAAG	AATTAATGAAAGCTCCCTGCC	227bp	chr17:40491289-40491515
6	AACAGGGAGCCTTCTCTTGG	ATGACCAGGCTCCTTTGAGG	382bp	chr17:40490606-40490987
7	AAGGAGTTTTTTCTTCCTTCGC	AACCAAGCAAGTTCTGCCAC	203bp	chr17:40489697-40489899
8	GAAGTTCCTGCTCTGGAGTTG	AGTACCAATTCTGTGGGCCTG	324bp	chr17:40489368-40489691
9	GTGGTCTCCATGTCTTCAGTATTTC	CCCGCCTCTCACTCTACCAC	327bp	chr17:40485835-40486161
10	TCACGTGGTAGAGTGAGAGGC	CACCTGGAAAGAATGACCCTG	264bp	chr17:40485595-40485858
11	GACAGCTTGGCCTATTTACCTG	TGACAATGCACCCCAAGGCT	327bp	chr17:40483340-40483666
12-13	AGTAAATAACAGGTGGTCAAAGTAGG	AAAGGCAGGTGTCCTGTGAG	385bp	chr17:40481490-40481874
14	CACTTGGTATGTGGGAGGAGC	GAACAGGTTGATGTTTCTAATTCTGG	252bp	chr17:40481326-40481577
15	TGCTGCTTAGACTGGTCTCG	CAGTAATCATTCCACCTTCTCTTG	309bp	chr17:40478034-40478342
16	ATGTCCTTTCATTCTGAGCCC	TGTTTAGATGAGGGATGGTGC	292bp	chr17:40476888-40477179
17	CCTAGTATAGACAATGAGCACCATCC	TCAGTAGACATGGCCCAAATG	286bp	chr17:40476640-40476925
18	CCATTGTGGGATGAATGTGTG	ACCAAGAGTTCAGGGCCTTC	242bp	chr17:40475498-40475739
19	GTAGACGGGCCTGAGGATTTG	ATTGCTAACAGGGCATCCATC	279bp	chr17:40475192-40475470
20	TGCACAGATGTAACCAAGTCC	TGCTTGCAACTAGAAGCAGTG	311bp	chr17:40474950-40475260
21	GAACAGGGTGTTCAGGGTCTC	CCCAGGTTATTCAGGCATTTG	447bp	chr17:40474201-40474647
22	GCTTCCTCCAGCTCTGCTTAC	GTCCCATTCCCAGGGATAAC	179bp	chr17:40469133-40469311
23	TTCCATTGTGTCTTGTCAACC	CCAACTTCCCTTATAGGGACAAAG	215bp	chr17:40468763-40468977
24a	TGGATAGCAGTGGTAGCCTGG	GATCACCCACATTCACTCATTTC	360bp	chr17:40467566-40467925

IV. Ethanol precipitation

V. Sequencing analysis

2xGC Buffer 2	5 µL
2.5mM dNTP	1.6 µL
5pmol/µL Fw primer	1 µL
5pmol/µL Rev primer	1 µL
LA Taq	0.1 µL
Genomiphi product	0.5 µL
Sterilized distilled water	0.8 µL

2xGC Buffer 1	5 µL
2.5mM dNTP	1.6 µL
5pmol/µL Fw primer	1 µL
5pmol/µL Rev primer	1 µL
LA Taq	0.1 µL
Genomiphi product	0.5 µL
Sterilized distilled water	0.8 µL

5xPrimeSTAR GXL Buffer	2 µL
2.5mM dNTP	0.8 µL
5pmol/µL Fw primer	1 µL
5pmol/µL Rev primer	1 µL
PrimeSTAR GXL DNA Polymerase	0.2 µL
Genomiphi product	0.5 µL
Sterilized distilled water	4.5 µL