SCHDGP005 PCR report

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SCHDGP005 PCR report 2010.02.23 1. Primer list Primer set Name Sequence Size (bp) 1 hSOST-e1-F GAGGGAAACATGGGACCAG 500 hSOST-e1-R ACTGTTCCTCGACCAGTGCT 2 hSOST-e2-1-F CAAAAGCCTCTGGAGACAGG 441 hSOST-e2-1-R AAGTCCTTGAGCTCCGACTG 3 hSOST-e2-2-F GTGGTGGCGACCTACTGG 420 hSOST-e2-2-R AGGGCCTGGAAGGTCTCAG Primer set Name Sequence Size (bp) 4 hSOST-1F tctctgggtcacctgggagt 1065 hSOST-1R cagggctatcttgcagttgg 5 hSOST-2F ctcgaaggggaagaaacagg 1021 hSOST-2R cccactctgaagagcaagca 6 hSOST-3F cccctgaccacacacacatt 1044 hSOST-3R ccattacacacgcaatgcag 7 hSOST-4F aaggaatttgggagaagcca 1012 hSOST-4R agaggacagaaatgtggggc

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2010.02.23. SCHDGP005 PCR report. 1. Primer list. 2010.02.23. SCHDGP005 PCR report. 2. The Result of PCR. 2-2. PCR Condition. 2-1. PCR reaction combination. 2010.02.23. SCHDGP005 PCR report. 2-4. Electrophoresis : primer set 1,2,3 _ 추가 PCR. 2-3. Electrophoresis. 1kb Ladder. - PowerPoint PPT Presentation

Transcript of SCHDGP005 PCR report

Page 1: SCHDGP005 PCR report

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

Page 2: SCHDGP005 PCR report

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

Page 3: SCHDGP005 PCR report

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

Page 4: SCHDGP005 PCR report

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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