SCHDGP005 PCR report 2010.02.23
1. Primer list
Primer set Name Sequence Size (bp)
1hSOST-e1-F GAGGGAAACATGGGACCAG
500hSOST-e1-R ACTGTTCCTCGACCAGTGCT
2hSOST-e2-1-F CAAAAGCCTCTGGAGACAGG
441hSOST-e2-1-R AAGTCCTTGAGCTCCGACTG
3hSOST-e2-2-F GTGGTGGCGACCTACTGG
420hSOST-e2-2-R AGGGCCTGGAAGGTCTCAG
Primer set Name Sequence Size (bp)
4hSOST-1F tctctgggtcacctgggagt
1065hSOST-1R cagggctatcttgcagttgg
5hSOST-2F ctcgaaggggaagaaacagg
1021hSOST-2R cccactctgaagagcaagca
6hSOST-3F cccctgaccacacacacatt
1044hSOST-3R ccattacacacgcaatgcag
7hSOST-4F aaggaatttgggagaagcca
1012hSOST-4R agaggacagaaatgtggggc
SCHDGP005 PCR report
2. The Result of PCR
2-1. PCR reaction combination 2-2. PCR Condition
Temp. Time CyclesFinal Reaction
vol.
95 ℃ 10 min 1
50ul
Ex-Taq
94 ℃ 30 sec
3555 ℃ 30 sec
72 ℃ 30 sec
72℃ 10 min 1
4℃ Storage 1
PCR reaction
SOST 체코 gDNA (50ng) x ul
10x Ex Taq buffer(TaKaRa) 5 ul
25mM dNTP(TaKaRa) 5 ul
Ex Taq(TaKaRa) 0.5ul
5x Band Doctor 10ul
Forward primer/Reverse Primer set (10pmol) 1 ul
Distilled water 28.5-x ul
Total 50ul
2010.02.23
SCHDGP005 PCR report
1kb Ladder
1.5% agarose gel 1.5% agarose gel
2-3. Electrophoresis 2-4. Electrophoresis : primer set 1,2,3 _ 추가 PCR
3. Sequencing : Primer set 1, 4~7 PCR product 를 gel elution 하여 양방향 sequencing
1 2 3 4 5 6 7
Primer set1 2 3
Primer set
2010.02.23
SCHDGP005 PCR report
4. PCR 추가 : template 농도 변경
4-1. PCR reaction combination
4-2. PCR Condition ; 이전과 동일
PCR reaction
SOST 체코 gDNA (100ng) x ul
10x Ex Taq buffer(TaKaRa) 5 ul
25mM dNTP(TaKaRa) 5 ul
Ex Taq(TaKaRa) 0.5ul
5x Band Doctor 10ul
Forward primer/Reverse Primer set (10pmol) 1 ul
Distilled water 28.5-x ul
Total 50ul
2010.02.23
4-3. Electrophoresis
1% agarose gel
1 2 3
Primer set
5. Sequencing : 2,3 번 primer set 는 sequencing 못함
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