RAPID Lite

PTPRC

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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
1-2	GTGCAATGCCATTCTACTGAC	CAATCTTGATGGTTTACAGCAG	528bp	chr1:198608075+198608602
3	GCCACTTGGTGAATGTTCTATC	CGTATAAGCAGAAAGTTTAAGAACAAG	310bp	chr1:198661359+198661668
4	GAGCATACATTTAGGGTATGATTCAC	ATACTAATTTCCTGCAATTTCTGCAC	412bp	chr1:198665774+198666185
5	AGGCTATGTAAATAGAGGACTCA	CTGTAGTCTCACAATTCTCACTATTA	552bp	chr1:198668494+198669045
6	TAGAATGCATGTGTCGTGGTA	TTTGGTGTTGATACGTGAACA	432bp	chr1:198671369+198671800
7	ATGTCAAATCAAACGAGGACT	CTTTCTGATGAATTATGACAGTAAC	426bp	chr1:198672298+198672723
8	CCGCCAATTAGATGCGTACC	CCTCCATCATCGCTCTTGTAT	518bp	chr1:198673329+198673846
9	AAATGATATGGAGGCACCTAA	CAGACTAGCTCCTGTGACCAC	453bp	chr1:198675718+198676170
10	CTTGCCAAATTCTATGTCATTAG	CATATCCCTAGATGCACTTGC	761bp	chr1:198677138+198677898
11	TTGAATAAGGGAAACTCAGCCTGTA	TGCATGACTTGGCAATGGACT	554bp	chr1:198678613+198679166
12	CAAATTAAATGAGCCCAGATGC	GGCTGGAAAGCTTTGAATGAG	443bp	chr1:198681928+198682370
13	GGTAGTACTGACTTTTAACGACTTAC	ACAGAGGTTAGTAGGCTAATGC	599bp	chr1:198685759+198686357
14	GGGATTGCAATAACTCTCATGGCT	CTCAATCTCATGTGAGCAGTG	551bp	chr1:198686982+198687532
15	CCTTGAAGTTTAAAGCAAATCAGCA	TTGTATCTAAGGCCCACTTCTGCTA	596bp	chr1:198691395+198691990
16	AGCAAGTTATTTGACAATCGT	AGATGTGGTTGTTTCTGACCA	531bp	chr1:198697419+198697949
17	CTCTGCTCAAGGCTCTTCCTC	GGCTGCAATAAGCAGAGATTG	367bp	chr1:198698087+198698453
18	TTCTCAGATGAAATGTGTACGAG	CATTAAAATATTTACTAGCTCTACACT	449bp	chr1:198700617+198701065
19-20	AGGATATCTCATTTACTCAAAACG	GTCCTCACCTATACGGCACAT	691bp	chr1:198701337+198702027
21-22	TGAAGCAATTCCTCTAACTCAC	GAAACAGGAATCAAGTTAAGAGTC	573bp	chr1:198703236+198703808
23	CTGACAACTGATTATGTCCTCTC	GAGTTGGACAAAAACGTCTTG	595bp	chr1:198703986+198704580
24	TTGAGTAGTTTATGTTTACACGAG	TCAATTCCGATATAGGTTCCT	518bp	chr1:198710882+198711399
25	TTAGAGACATCTTTCCCTTTGGC	GGAAATCATCTACTGTTTGAGGTTG	396bp	chr1:198711226+198711621
26	TTGCCATGAATTGTTCCAATATG	GCTCCTTTGGTTCTGGTGAAG	392bp	chr1:198713066+198713457
27	TCTTTAGTGCATACTAGCCCTGTC	CAAAGCCCATTGTAAATGTTCAG	504bp	chr1:198717024+198717527
28	TGTTGTTTGACATGGAATTATGTTC	GCTTAATGAATGGCCCTAATAAC	451bp	chr1:198718306+198718756
29	TACTGAGCTCTCTTCTGTAGTAACG	TAGCTGTTATTTGCTGTGACG	463bp	chr1:198719568+198720030
30	CCAGTCACCTCGATTGTGCTC	TTATATGATGGCCTTGCCTGC	465bp	chr1:198721132+198721596
31	CTATGCAGGCAAGGCCATCA	CCAAGAGAGTGACCGACTCCA	470bp	chr1:198721572+198722041
32	GGTTTCCTTATCTATCATACTGGCA	GCTCTTCATGAACTGTCACAAC	726bp	chr1:198723057+198723782
33a	TGCAGTCAACCCTTTAGCACCA	CGTCTGTACAAACTGTTCTGGCAA	655bp	chr1:198724845+198725499

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
1-2	GTGCAATGCCATTCTACTGAC	CAATCTTGATGGTTTACAGCAG	528bp	chr1:198608075+198608602
3	GCCACTTGGTGAATGTTCTATC	CGTATAAGCAGAAAGTTTAAGAACAAG	310bp	chr1:198661359+198661668
4	GAGCATACATTTAGGGTATGATTCAC	ATACTAATTTCCTGCAATTTCTGCAC	412bp	chr1:198665774+198666185
5	AGGCTATGTAAATAGAGGACTCA	CTGTAGTCTCACAATTCTCACTATTA	552bp	chr1:198668494+198669045
6	TAGAATGCATGTGTCGTGGTA	TTTGGTGTTGATACGTGAACA	432bp	chr1:198671369+198671800
7	ATGTCAAATCAAACGAGGACT	CTTTCTGATGAATTATGACAGTAAC	426bp	chr1:198672298+198672723
8	CCGCCAATTAGATGCGTACC	CCTCCATCATCGCTCTTGTAT	518bp	chr1:198673329+198673846
9	AAATGATATGGAGGCACCTAA	CAGACTAGCTCCTGTGACCAC	453bp	chr1:198675718+198676170
10	CTTGCCAAATTCTATGTCATTAG	CATATCCCTAGATGCACTTGC	761bp	chr1:198677138+198677898
11	TTGAATAAGGGAAACTCAGCCTGTA	TGCATGACTTGGCAATGGACT	554bp	chr1:198678613+198679166
12	CAAATTAAATGAGCCCAGATGC	GCAATTCTAAGATCAGGGAGT	417bp	chr1:198681928+198682344
13	GGTAGTACTGACTTTTAACGACTTAC	ACAGAGGTTAGTAGGCTAATGC	599bp	chr1:198685759+198686357
14	GGGATTGCAATAACTCTCATGGCT	CTCAATCTCATGTGAGCAGTG	551bp	chr1:198686982+198687532
15	CCTTGAAGTTTAAAGCAAATCAGCA	TTGTATCTAAGGCCCACTTCTGCTA	596bp	chr1:198691395+198691990
16	AGCAAGTTATTTGACAATCGT	AGATGTGGTTGTTTCTGACCA	531bp	chr1:198697419+198697949
17	CTCTGCTCAAGGCTCTTCCTC	GGCTGCAATAAGCAGAGATTG	367bp	chr1:198698087+198698453
18	TTCTCAGATGAAATGTGTACGAG	CATTAAAATATTTACTAGCTCTACACT	449bp	chr1:198700617+198701065
19-20	AGGATATCTCATTTACTCAAAACG	GTCCTCACCTATACGGCACAT	691bp	chr1:198701337+198702027
21-22	TGAAGCAATTCCTCTAACTCAC	GAAACAGGAATCAAGTTAAGAGTC	573bp	chr1:198703236+198703808
23	CTGACAACTGATTATGTCCTCTC	GAGTTGGACAAAAACGTCTTG	595bp	chr1:198703986+198704580
24	TTGAGTAGTTTATGTTTACACGAG	TTTTTTTTTGTTCGTTTTTC	298bp	chr1:198710882+198711179
25	TTAGAGACATCTTTCCCTTTGGC	GGAAATCATCTACTGTTTGAGGTTG	396bp	chr1:198711226+198711621
26	TTGCCATGAATTGTTCCAATATG	GCTCCTTTGGTTCTGGTGAAG	392bp	chr1:198713066+198713457
27	TCTTTAGTGCATACTAGCCCTGTC	CAAAGCCCATTGTAAATGTTCAG	504bp	chr1:198717024+198717527
28	TGTTGTTTGACATGGAATTATGTTC	GCTTAATGAATGGCCCTAATAAC	451bp	chr1:198718306+198718756
29	TACTGAGCTCTCTTCTGTAGTAACG	TAGCTGTTATTTGCTGTGACG	463bp	chr1:198719568+198720030
30	CCAGTCACCTCGATTGTGCTC	TTATATGATGGCCTTGCCTGC	465bp	chr1:198721132+198721596
31	CTATGCAGGCAAGGCCATCA	CCAAGAGAGTGACCGACTCCA	470bp	chr1:198721572+198722041
32	GGTTTCCTTATCTATCATACTGGCA	GAAAAAAAAAATTCCGAGCA	526bp	chr1:198723057+198723582
33a	TGCAGTCAACCCTTTAGCACCA	CGTCTGTACAAACTGTTCTGGCAA	655bp	chr1:198724845+198725499

IV. Ethanol precipitation

V. Sequencing analysis

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

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