Elution of DNA by Crush and Soak Method
- 최초 등록일
- 2014.09.13
- 최종 저작일
- 2014.08
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한컴오피스
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목차
1. < INTRODUCTION >
2. < Materials >
3. < Method >
4. < References >
본문내용
1. Polyacylamide gel elution
① crush and soak method
- gel을 부수고 elution buffer에 적시어 diffusion으로 DNA elution.
② electroelution
- membrane에 gel slice를 넣고 전기를 걸어 DNA를 elution하는 방법.
③ Protoprep (glass powder)
2. Crush and soak method
: migration depending on size (PAGE) and elution of the DNA using elusion buffer.
- recover DNA from polyacrylamide gels
- modification of the technique described by Maxam and Gilbert (1977, 1980)
* DNA sequencing - chemical sequencing by Maxam and Gilbert (1977) (별지1)
① ds DNA labelling(32P)
② base alteration
③ ds DNA separation → two ss DNA strands(gel electrophoresis)
④ template strand purification
⑤ cleavage reaction
⑥ electrophoresis → DNA sequencing
- requires time but little labor
- recovery of < 30%~90% yield
- used to isolate double strand DNA (neutral polyacrylamide gel) and single strand DNA (denaturing polyacrylamide gel(urea)) ; suitable for use as a hybridzation probe, as a probe in gel-retention assays, or as a template in chemical sequencing and enzymatic reactions.
참고 자료
Sambrook and Russell. 2001. Molecular cloning 3rd. 5.51-54/10.11-16/12.74-82
http://nationaldiagnostics.com/article_info.php/articles_id/40
http://www.protocol-online.org/prot/Molecular_Biology/DNA/DNA_Extraction___ Purification/DNA_Extraction_from_Acrylamide_Gel/index.html
http://www.wmin.ac.uk/~redwayk/lectures/sequence
