JP2005013122A5 - - Google Patents
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- JP2005013122A5 JP2005013122A5 JP2003183694A JP2003183694A JP2005013122A5 JP 2005013122 A5 JP2005013122 A5 JP 2005013122A5 JP 2003183694 A JP2003183694 A JP 2003183694A JP 2003183694 A JP2003183694 A JP 2003183694A JP 2005013122 A5 JP2005013122 A5 JP 2005013122A5
- Authority
- JP
- Japan
- Prior art keywords
- oligonucleotide
- primer
- chimeric
- value
- chimeric oligonucleotide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
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Description
【特許請求の範囲】
【請求項１】
遺伝子増幅試薬の安定化方法であって、遺伝子増幅反応に使用するキメラオリゴヌクレ
オチドプライマーと相補的な塩基配列を有するオリゴヌクレオチドを少なくとも一つ、遺
伝子増幅反応液中に共存させることを特徴とする方法であって、当該オリゴヌクレオチド
がＤＮＡ又はＲＮＡオリゴヌクレオチド、あるいはＤＮＡ−ＲＮＡキメラオリゴヌクレオ
チドから選択され、かつ当該オリゴヌクレオチドと前記キメラオリゴヌクレオチドプライ
マーの間のＴｍ値に対する、前記キメラオリゴヌクレオチドプライマーと該プライマーの
塩基配列と相補的な標的遺伝子の核酸配列との間のＴｍ値の比が、１．２〜３．３の範囲
であり、さらに当該オリゴヌクレオチドの３’末端が伸長できないようにブロックされた
オリゴヌクレオチドであることを特徴とする方法。
【請求項２】
請求項１記載の方法のための増幅試薬ミクスチャーであって、
（１）キメラオリゴヌクレオチドプライマー、
（２）（１）のキメラオリゴヌクレオチドプライマーと相補的な塩基配列を有するオリゴ
ヌクレオチドであって、当該オリゴヌクレオチドと前記キメラオリゴヌクレオチドプライ
マーの間のＴｍ値に対する、前記キメラオリゴヌクレオチドプライマーと該プライマーの
塩基配列と相補的な標的遺伝子の核酸配列との間のＴｍ値の比が、１．２〜３．３の範囲
であり、さらに当該オリゴヌクレオチドの３’末端が伸長できないようにブロックされた
オリゴヌクレオチド、
（３）鎖置換活性を有するＤＮＡポリメラーゼ
（４）ＲＮａｓｅＨ、及び
（５）反応緩衝液
（６）ｄＮＴＰｓ
からなることを特徴とするミクスチャー。
[Claims]
[Claim 1]
A method for stabilizing a gene amplification reagent, wherein at least one oligonucleotide having a base sequence complementary to a chimeric oligonucleotide primer used in a gene amplification reaction is allowed to coexist in the gene amplification reaction solution Said chimeric oligonucleotide primer and said primer, wherein said oligonucleotide is selected from DNA or RNA oligonucleotide or DNA-RNA chimeric oligonucleotide and said Tm value between said oligonucleotide and said chimeric oligonucleotide primer The Tm value ratio between the nucleotide sequence of the target and the complementary target gene nucleic acid sequence was in the range of 1.2 to 3.3, and the 3 ′ end of the oligonucleotide was blocked so that it could not be extended. Be an oligonucleotide Feature method.
[Claim 2]
An amplification reagent mixture for the method of claim 1, comprising:
(1) a chimeric oligonucleotide primer,
(2) An oligonucleotide having a base sequence complementary to the chimeric oligonucleotide primer of (1), wherein the chimeric oligonucleotide primer and the primer have a Tm value between the oligonucleotide and the chimeric oligonucleotide primer. The ratio of the Tm value between the nucleotide sequence and the complementary target gene nucleic acid sequence is in the range of 1.2 to 3.3, and the oligonucleotide is blocked so that the 3 ′ end of the oligonucleotide cannot be extended. nucleotide,
(3) DNA polymerase having strand displacement activity (4) RNase H, and (5) reaction buffer (6) dNTPs
A mixture characterized by consisting of
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2003183694A JP2005013122A (en) | 2003-06-27 | 2003-06-27 | Method for stabilizing reagent for amplifying gene |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2003183694A JP2005013122A (en) | 2003-06-27 | 2003-06-27 | Method for stabilizing reagent for amplifying gene |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2005013122A JP2005013122A (en) | 2005-01-20 |
| JP2005013122A5 true JP2005013122A5 (en) | 2006-07-20 |
Family
ID=34183666
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2003183694A Pending JP2005013122A (en) | 2003-06-27 | 2003-06-27 | Method for stabilizing reagent for amplifying gene |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2005013122A (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP4960536B2 (en) * | 2010-02-25 | 2012-06-27 | パナソニック株式会社 | Method for amplifying a target sequence in double-stranded DNA |
-
2003
- 2003-06-27 JP JP2003183694A patent/JP2005013122A/en active Pending
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