QIAGEN Plasmid Kits for Plasmid DNA Extraction

10 mgまでのトランスフェクショングレードのプラスミドおよびコスミドDNA精製用

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QIAGEN Plasmid Mini Kit (100)

カタログ番号 / ID. 12125

100 QIAGEN-tip 20, Reagents, Buffers

CA$1,084.00

ログインアカウントの価格を確認するには

KitBufferColumnCartridge

Plasmid Kit

Plasmid Buffer Set

QIAGEN-tip

QIAfilter Cartridge

カラムタイプ

Mini

Midi

Maxi

Mega

Giga

調製

100

数量

プラスミドDNA抽出用QIAGEN Plasmid Kitsは分子生物学的アプリケーション用であり、疾病の診断、予防、あるいは治療に使用することはできません。

✓ オンライン注文による24時間年中無休の自動処理システム

✓ 知識豊富で専門的な製品&テクニカルサポート

✓ 迅速で信頼性の高い（再）注文

特徴

CsCl密度勾配遠心法を2回行なって得られる純度に匹敵
高収量のプラスミドDNA
経済的な調製法
LyseBlue試薬により最適な溶解操作と最大のDNA収量

製品詳細

QIAGEN Plasmid Kits はオープンカラム方式の陰イオン交換チップを用いてトランスフェクショングレードのプラスミドDNA 精製を実現します。ライセート清澄化およびイソプロパノール沈殿は遠心操作により行ないます。

パフォーマンス

QIAGEN Plasmid Kitsでは、プラスミドDNAを効率的に精製するためのオープンカラム方式のQIAGEN陰イオン交換チップを使用しています。それぞれ、10 mg（Giga）、2.5 mg（Mega）、500 µg（Maxi）、100 µg（Midi）、20 µg (Mini) までのトランスフェクショングレードの高コピー数プラスミドDNAを培養液から精製します（培養液の容量はプラスミドコピー数、挿入DNAのサイズ、宿主株、培養液に依存）。QIAGEN Plasmid Kitsで精製したプラスミドDNAは、トランスフェクション（図""）、クローニング、in vitro 転写などのアプリケーションで使用するのに最適です。

図参照

Transfection efficiency versus plasmid purification method.

原理

QIAGEN-tip 中の非常にユニークな陰イオン交換樹脂は核酸精製を目的として開発されました。本製品の優れた核酸分離能力により、CsCl 密度勾配遠心分離法を2回行なって得たDNAの純度に匹敵、あるいはそれ以上の純度のDNAが調製されます。充填済みQIAGEN-tipsはオープンカラムで操作し、乾燥することはなく、プラスミド調製に必要なマニュアルでの作業時間を短縮できます。全てのQIAGENプラスミド精製システムでは、ユーザーおよび環境への影響が最小限となるように、フェノール、クロロホルム、臭化エチジウム、CsCl等の有害な試薬を一切使用していません。

製品仕様
特徴	Plasmid Giga Kit	Plasmid Mega Kit	Plasmid Maxi Kit	Plasmid Midi Kit	Plasmid Mini Kit
Applications	Transfection, cloning, sequencing, capillary sequencing, etc.	Transfection, cloning, sequencing, capillary sequencing, etc.	Transfection, cloning, sequencing, capillary sequencing, etc.	Transfection, cloning, sequencing, capillary sequencing, etc.	Transfection, cloning, sequencing, capillary sequencing, etc.
Culture volume/starting material	2.5–5 liters culture volume	500 ml – 2.5 liters culture volume	100–500 ml culture volume	25–100 ml culture volume	3–10 ml culture volume
Elution volume	Variable	Variable	Variable	Variable	Variable
Plasmid type	High-copy, low-copy, cosmid DNA	High-copy, low-copy, cosmid DNA	High-copy, low-copy, cosmid DNA	High-copy, low-copy, cosmid DNA	High-copy, low-copy, cosmid DNA
Processing	Manual (centrifugation)	Manual (centrifugation)	Manual (centrifugation)	Manual (centrifugation)	Manual (centrifugation)
Sample per run	1 sample per run	1 sample per run	1 sample per run	1 sample per run	1 sample per run
Technology	Anion-exchange technology	Anion-exchange technology	Anion-exchange technology	Anion-exchange technology	Anion-exchange technology
Time per run	320 min	220 min	160 min	150 min	80 min
Yield	<10 mg	<2.5 mg	<500 µg	up to 100 µg	<20 µg

操作手順

QIAGEN Plasmid Kitsを使用して、遠心操作でバクテリアライセートを清澄化します。清澄化したライセートを陰イオン交換チップ上にアプライすると、適切な低塩あるいはpH条件でプラスミドDNAが選択的に結合します。RNA、タンパク質、二次代謝物、その他の低分子量不純物が、中濃度の塩による洗浄で取り除かれ、高純度プラスミドDNAが高塩濃度のバッファーで溶出されます（フローチャート" QIAGEN Plasmid Kits 操作の比較"参照）。イソプロパノール沈殿によりDNAが濃縮・脱塩され、遠心操作により回収されます。

図参照

QIAGEN Plasmid Kit procedures.

アプリケーション

QIAGEN Plasmid Kits を用いて精製したプラスミドDNAは、以下のようなアプリケーションに最適です。

トランスフェクション
クローニング
PCR
In vitro転写

Example 1
	12123	12125	12143	12145	12145X4	12162	12163	12165
Product	Plasmit Kit	Plasmit Kit	Plasmit Kit	Plasmit Kit	Plasmit Kit	Plasmit Kit	Plasmit Kit	Plasmit Kit
Product Type	Kit	Kit	Kit	Kit	Kit	Kit	Kit	Kit
Column Type	Mini	Mini	MIDI	MIDI	MIDI	MAXI	MAXI	MAXI
Preparations	25	100	25	100	400	10	25	100

Example 2
CatNo	Product	Product Type	Column Type	Preparations
12123	Plasmit Kit	Kit	Mini	25
12125	Plasmit Kit	Kit	Mini	100
12143	Plasmit Kit	Kit	MIDI	25
12145	Plasmit Kit	Kit	MIDI	100
12145X4	Plasmit Kit	Kit	MIDI	400
12162	Plasmit Kit	Kit	MAXI	10
12163	Plasmit Kit	Kit	MAXI	25
12165	Plasmit Kit	Kit	MAXI	100

裏付けデータと数値

Transfection efficiency versus plasmid purification method.

Different pRSVcat DNA preparations using the methods indicated were introduced into the indicated cell lines by liposome-mediated transfection, and the efficiencies determined by measuring CAT expression levels after 40 h. Each bar represents the mean of 4 independent transfections (2 transfections with each of 2 independent plasmid preparations). The highest transfection efficiency was achieved with QIAGEN Plasmid Kits.

仕様

特徴	仕様
technology	Anion-exchange technology
culturevolumestartingmaterial	3 ml–5 liters culture volume
yield	<20 µg to <10 mg
processing	Manual (gravity flow)
samplesperrunthroughput	1 sample per run
timeperrunorprepperrun	80–320 min
applications	Transfection, cloning, sequencing, capillary sequencing etc.
plasmidtype	High-copy, low-copy, cosmid DNA

リソース

April 2012

This protocol is designed for the rapid, easy, and non-toxic preparation of up to 2 mg genomic DNA from not more than 2 g of tissue using QIAGEN-tip 2500. QIAGEN® Genomic-tips 20/G, 100/G, and 500/G can also be used with this protocol by reducing the amount of starting material according to the table on page 2. The purified genomic DNA ranges in size from 50-150 kb.

Endotoxin-free DNA is essential for gene therapy research and will improve transfection into sensitive eukaryotic cells.

This protocol is for purification of up to 100 µg endotoxin-free plasmid DNA using QIAGEN-tip 100.

This protocol is designed to provide up to 150 μg BAC/PAC/P1 DNA or up to 400 μg cosmid DNA.

This protocol is designed for isolation of up to 200 μg RNA from 150 mg plant tissue or up to 1 mg RNA from 600 mg plant tissue and is for use with QIAGEN-tip 100 or QIAGEN-tip 500, respectively.

This procedure has been used successfully for isolation of 150-250 kb BAC DNA from a mouse-BAC library cloned in pBeloBAC11 from Escherichia coli strain HB101/r. The yield of BAC DNA from 100 ml culture was typically 20-40 μg.

The procedure has been used successfully for isolation of high- and low-copy-number plasmids from various Bacillus subtilis strains. Yield of plasmid DNA was typically 10-20 µg plasmid DNA from 100 ml culture.

The procedure has been used successfully for isolation of 110 kb P1 DNA (pAdsacBII with an 80 kb insert) from Escherichia coli strain NS3529. Yield of P1 DNA was typically 10-50 µg from 500 ml culture.

The procedure has been used successfully for isolation of linear plasmids from Borrelia burgdorferi sensu lato species, which include Borrelia burgdorferi sensu stricto, Borrelia afzelli, and Borrelia garinii.

The procedure has been used successfully for isolation of high-copy-number plasmids from Citrobacter freundii. Yield of plasmid DNA was typically 3-8 µg DNA per ml culture.

The procedure has been used successfully for isolation of different medium-copy-number plasmids carrying pHM1519 or pBL1 origins of replication from Corynebacterium glutamicum ATCC 13032. Yield of plasmid DNA was typically 0.4-1.5 µg per ml LB culture, although yield was dependent on the vector, the insert, and the size of the plasmid.

The procedure has been used successfully for isolation of cryptic plasmids (pLC2-based) from mesophilic Lactobacillus strains such as L. sake and L. curvatus. Yield of plasmid DNA was typically 10-20 µg plasmid DNA from 100 ml culture.

The procedure has been used successfully for isolation of the large (128 kb), very-low-copynumber (1-2 copies per cell) plasmid pHCG3 and its derivatives from Oligotropha carboxidovorans. Yield of plasmid DNA was typically 3-6 µg plasmid DNA from 200 ml culture.

The procedure has been used successfully for isolation of high-copy-number plasmids from Proteus vulgaris and Proteus mirabilis. Yield of plasmid DNA was typically 3-8 µg DNA per ml culture.

The procedure has been used successfully for isolation of a variety of medium-copy-number shuttle vectors from S. xylosus, S. carnosus, S. epidermidis, and S. aureus. Yield of plasmid DNA was typically 2-10 µg from 50 ml culture.

The procedure has been used successfully for isolation of plasmids SCP2 and SCP2* as well as the plasmids listed in Table 1 (see next page) from S. coelciolor and S. lividans strains.

Download Safety Data Sheets for QIAGEN product components.

April 2012

出版物

Oncocytic change in pleomorphic adenoma: molecular evidence in support of an origin in neoplastic cells.

Di Palma S; Lambros MB; Savage K; Jones C; Mackay A; Dexter T; Iravani M; Fenwick K; Ashworth A; Reis-Filho JS;

J Clin Pathol; 2006; 60 (5):492-9 2006 Feb 7 PMID:16467165

High-level carbapenem resistance in a Klebsiella pneumoniae clinical isolate is due to the combination of bla(ACT-1) beta-lactamase production, porin OmpK35/36 insertional inactivation, and down-regulation of the phosphate transport porin phoe.

Kaczmarek FM; Dib-Hajj F; Shang W; Gootz TD;

Antimicrob Agents Chemother; 2006; 50 (10):3396-406 2006 Oct PMID:17005822

Haploinsufficiency of C2GnT-I glycosyltransferase renders T lymphoma cells resistant to cell death.

Cabrera PV; Amano M; Mitoma J; Chan J; Said J; Fukuda M; Baum LG;

Blood; 2006; 108 (7):2399-406 2006 Jun 15 PMID:16778138

RepAM of the Amycolatopsis methanolica integrative element pMEA300 belongs to a novel class of replication initiator proteins.

Te Poele EM; Kloosterman H; Hessels GI; Bolhuis H; Dijkhuizen L;

Microbiology (Reading); 2006; 152 (Pt 10):2943-2950 2006 Oct PMID:17005975

Involvement of Bcl-X(L) deamidation in E1A-mediated cisplatin sensitization of ovarian cancer cells.

Chang CY; Lin YM; Lee WP; Hsu HH; Chen EI;

Oncogene; 2006; 25 (18):2656-65 2006 Apr 27 PMID:16331250

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QIAGEN Plasmid Kits for Plasmid DNA Extraction

10 mgまでのトランスフェクショングレードのプラスミドおよびコスミドDNA精製用

QIAGEN Plasmid Mini Kit (100)

特徴

製品詳細

パフォーマンス

図参照

原理

操作手順

図参照

アプリケーション

裏付けデータと数値

Transfection efficiency versus plasmid purification method.

仕様

リソース

出版物

Oncocytic change in pleomorphic adenoma: molecular evidence in support of an origin in neoplastic cells.

High-level carbapenem resistance in a Klebsiella pneumoniae clinical isolate is due to the combination of bla(ACT-1) beta-lactamase production, porin OmpK35/36 insertional inactivation, and down-regulation of the phosphate transport porin phoe.

Haploinsufficiency of C2GnT-I glycosyltransferase renders T lymphoma cells resistant to cell death.

RepAM of the Amycolatopsis methanolica integrative element pMEA300 belongs to a novel class of replication initiator proteins.

Involvement of Bcl-X(L) deamidation in E1A-mediated cisplatin sensitization of ovarian cancer cells.