TaKaRa 1kbmarker使用说明书
TAKARA 实验室常规试剂配制方法
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<AHIm=CODklPi]im:;031*-2./4,+586779\_V_[_PUY\RQTXYWYS^QO\OWYSYXWUXR:bkkiNIIlllHk_d_j_H`hfH`gOfic`ceecg EKJJfaIfeFrqq 74p 76z 7:5256658vq 76ro Jf v 4z 7:5256658-}vq 76eyU .Aso ]q uq 76icuEKRZXoaYEQp ,vq 76A to |qoss 17hkWhLcAuo xSR /C r 76p SDYE nsq BGMAUZ\SELMfaIfeFsu 74p 76*./}vq 76ro Jf ros 4*./}-}vq 76eyU .Aso ]q uq 76icuEKRZXoaYEQp ,vq 76A to |qoss 17hkWhLcAuo xSR /C r 76p SDYE nsq BGMA]GS_eELJfaIfeFnsqBIVGMAr +ro Jlwgt\E -}r +rH .Aso ]q~yqq 76Nme 0uEKR_FoaA to O]v ,,1-~C~qos 76@EP`:~+,xoq A uo ]me 0u^j{[Qp ,r +A vo TvTzicYEdn ,svAwo ]q r 76z 7:5256658C rqq 74p 76DYb *ZXA xo u BGMA031*-2./4,+59678:UWPWTW JOSULKNRSQSMV KIUIQSMSRQORL;Y``^HGGaaaF`WZW_WFX][FX\i xif ]wZjnOFp>hfio y wv j |{l @hfoj y w j v|{l ?ihh 21<aU jfki /wv j |{l M ijfml /w j v|{l r qh 21I_Xyix@TAaUN@R_Xy iJbu ihh 21@LkLm[V<jf D^lYgQ@vr i x eCx<kf Rcs phh 21I_Xyi@EKTAaU<lf R_XyiS\oPJbu i x N@LkLm[V<mf HapW`u nh =qlf@Rc ihh 21Idh[VZjnOFp<nf l =BG<i xif ]w jmh 2y ws 1ap<t qh 21I_XyixaU ifpn /ws 1N@Jbu ihh 21<jf ]w j y y /s 1j ap<t qh 21_XyixaU iq /y /s 1j N@Jbu ihh 21@LkLm[V<kf D^lYgQ@vri x eCx<mf HapW`u nh =qlf@Rc ihh 21Idh[VZjnOFpM i 21I r 250-01103dihh 2/g 21e <031*-2./4,+59678:UWPWTW JOSULKNRSQSMV KIUIQSMSRQORL;Y``^HGGaaaF`WZW_WFX][FX\gff 21gd az g w bnmgs<V gn /bnmgu of 21f\|l{AFNgYRMhy gff 21At fdhh }2^cP^G[<hd p gff 21zyq `rT{UiG[J g w bnmgs h 21AZwSQ<id j >DH<jd Uiv nff 21Jf\|lAFNWCgY<kd ]e gf 21hkf 2w ur 1gsAUiIjE{<ld KU k x x +yt gs?v fdh 21;ALX5t xy mdf <md Uif\|lV`rTMhy g v <nd OoOq_[RA j >DH<od ktdUi k 21_[J h w w /r 1hgs<g v031*-2./4,+59678:UWPWTW JOSULKNRSQSMV KIUIQSMSRQORL;Y``^HGGaaaF`WZW_WFX][FX\x ,+w .Y rXRk q -9-4256p .2/D}]KHr H E P F 2I Uh `x ,q +w .Y r UO@x ,w .Y rXRk+0-9:r ;:8-.:Dw -M6S E P F 2I Uh `x ,+w .Y r UO@he I d[N.Y r /f m Pl [N.Y rF c i Q i X,{4Ft =T+D s k ^\l @ie i Q i X,{oF[?EL6W .Y rF Z d ;3C5e q 5eJ Vgp b DY B.Y r Q eniF VWK>>@je \.Y r~8g lg B mg C B Ft =9R R c Sj D <|AG _>F Z d ;3Vgp b@h vie t =a ogg 43^7*nF FVgvM :y@je `t l x x -yt D a gei 43>Fax7t fg neg @ke t 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RNA marker 1000
制品内容
RNA Marker RL1,000 6×Loading Buffer DEPC 处理水
20 μl 100 μl
1 ml
使用方法 A. 普通琼脂糖凝胶电泳时
1. 按下列组份配制 RNA Marker 样品。
RNA Marker RL1,000 6×Loading Buffer DEPC 处理水
RNA Marker RL1,000
使用说明书
Takara Code : D508A
包 装 量 : 约 20 次量
浓 度 : 约 200 ng/μl
保 存 : -80℃（短期保存可放置于-20℃）
制品说明 RNA Marker RL1,000是由体外转录得到的8条高纯度的单链RNA片 段组成的，其长度分别为1,000、800、600、500、400、300、200 和100 Bases，每微升本制品的RNA量约为200 ng。可用于RNA的 琼脂糖变性凝胶电泳（甲醛或乙二醛）或者普通的RNA琼脂糖凝胶 电泳等。每次取1 μl电泳时可使用约20次。
4. 甲醛变性琼脂糖凝胶配制方法如下： 加 6.0 g 高质量的琼脂糖到 93 ml DEPC 水中，煮沸直至完全溶 解，再加入适量的 DEPC 水定容至 93 ml。溶液冷却至 60℃左 右后，加入 30 ml 5×MOPS-EDTA Buffer 和 27 ml 37%甲醛（在 通风橱中操作），倒胶后于室温静置 1 小时。
1 μl 2 μl up to 10 μl
2. 均匀混合后 65℃加热 10 分钟，迅速冷却至室温（最好用 PCR 仪）。
3. 使用高质量的琼脂糖，用 1×TAE Buffer 制备 5%凝胶，制胶时 凝胶中请加入溴乙锭（终浓度：1 μg/ml）。
marker说明书
CERTIFICATE OF ANALYSISPageRuler ™Prestained Protein Ladder#SM067210 x 250 µl(for 100 mini gel applications 5 µl per well or 50 large gel applications 10 µl per well)Lot: Expiry Date:Storage: stable at 4°C for up to 3 months. For long term storage, store at -20°C.SM067_57_9.docDescriptionPageRuler ™Prestained Protein Ladder is a mixture of 10 recombinant, highly purified colored proteins with apparent molecular weights of 10 kDa to 170 kDa. Ladder proteins are covalently coupled with a blue dye except for two reference bands prestained with different colors. The 72 kDa reference band is orange and 10 kDa reference band is green.The ladder is supplied in gel loading buffer and isready-to-use: no heating, further dilution or addition of a reducing agent is required.ContentsApproximately 0.1-0.2 mg/ml of each protein in the storage buffer (62.5 mM Tris-H 3PO 4 (pH 7.5 at 25°C), 1 mM EDTA, 2% (w/v) SDS, 10 mM DTT, 1 mM NaN 3 and 33% (v/v) glycerol).Applications∙ Monitoring of protein separation during SDS-PAGE (1). ∙ Verifying Western transfer efficiency (2, 3).∙ Approximate sizing of proteins on SDS-polyacrylamidegels and Western blots.Instruction for Use❶ Thaw the ladder at room temperature for a few minutes to dissolve precipitated solids. DO NOT BOIL!❷ Mix gently, but thoroughly, to ensure the solution is homogeneous.❸ Load the following volumes of the ladder on an SDS-polyacrylamide gel:– 5 µl per well for mini gel,– 10 µl per well for large gel.Use the same volumes for Western blotting.❹ After the run is complete, stain the gel or perform Western transfer procedure as desired.Note•Each lot of the PageRuler™ Prestained Protein Ladder is calibrated against a precisely sized, PageRuler™ Unstained Protein Ladder and calculated apparent molecular weights are reported in the picture.•For precise molecular weight determinations use PageRuler™ Unstained Protein Ladder, #SM0661, see.•In 8 or 10% gels low molecular weight proteins may migrate with the dye front.•Loading volumes are intended for use in gels with a thickness of 0.75 mm. For thicker gels, the recommended loading volume should be increased.•PageRuler™ Prestained Protein Ladder could be used in Western blotting with all common membranes: PVDF, nylon and nitrocellulose.•Longer transfer times or higher transfer voltages may be required for Western blotting of large (>100 kDa) proteins.Lot specific MW, kDa4-20% Tris-glycine SDS-PAGEcontinued on back pageQUALITY CONTROL5 µl of PageRuler™ Prestained Protein Ladder resolves 10 bands of equal intensities in 4-20% SDS-PAGE (Tris-glycine buffer) and after Western blotting onto PVDF membrane.Quality authorized by: Jurgita Zilinskiene References1. Laemmli, U.K., Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature, 227, 680-685, 1970.2. Burnette, W.N., "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate – polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A, Anal. Biochem., 112 (2), 195-203, 1981.3. Towbin, H., et al., E lectrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications, Proc. Natl. Acad. Sci. USA, 76, 4350-4354, 1979.This product is manufactured under the license forStrep-tag® technology covered by US patents Nos.5,506,121, 6,103,493 and foreign counterparts. Related Products∙DualColor™ Protein Loading Buffer Pack #R1011∙Loading Buffer Pack #R0891∙Spectra™ Multicolor Broad Range Protein Ladder #SM1841∙PageRuler™ Unstained #SM0661∙PageRuler™ Plus Prestained Protein Ladder #SM1811∙PageSilver™ Silver Staining Kit #K0681∙PageBlue™ Protein Staining Solution #R0571∙10X Tris-glycine-SDS Buffer #B46∙10X Tris-tricine-SDS Buffer #B48∙DTT #R0861 ∙ProteoJET™ Mammalian Cell Lysis Reagent #K0301∙ProteoJET™ Cytoplasmic and Nuclear ProteinExtraction Kit #K0311∙Bradford Reagent, ready-to-use #R1271∙Bovine Serum Albumin Standard Set, ready-to-use #R1281∙Bovine Gamma Globulin Standard Set, ready-to-use #R1291PRODUCT USE LIMITATION.This product is developed, designed and sold exclusively for research purposes andin vitro use only. The product was not tested for use in diagnostics or for drugdevelopment, nor is it suitable for administration to humans or animals.Please refer to for Material Safety Data Sheet of the product.。
蛋白Marker说明书
蛋⽩Marker说明书PageRuler Prestained Protein LadderSM0671A prestained SDS-PAGE MW marker with contrasting colored reference bands at10 and 70kDa.The Thermo Scientific PageRuler Prestained Proteis a mixture of ten (10) blue-, orange- and green-srecombinant proteins (10 to 170kDa) for use as sizstandards in protein electrophoresis (SDS-PAGE)Western blotting.This prestained protein MW marker is designed fomonitoring the progress of SDS-polyacrylamide geelectrophoresis, for assessing transfer efficiency onylon and nitrocellulose membranes, and for estimapproximate size of separated proteins that have b visible with gel stains or Western blot detection reagents. The ladder contains one orange refer at 70kDa and one green band at 10kDa.Highlights:Size range– 10 proteins spanning 10 to 170kDaReady-to-use– supplied in a loading buffer for direct loading on gels; no need to boil Sharp bands– color-coded bands of similar intensity for easy visualizationQuality tested – each lot evaluated by SDS-PAGE and Western blottingTwo reference bands– orange at 70kDa and green at 10kDaMembrane-compatible– colored bands transfer to membranes for Western blotting Includes:Dye-stained proteins in 62.5mM Tris-H3PO4 (pH 7.5 at 25°C), 1mM EDTA, 2% SDS, 1 1mM NaN3 and 33% glycerol. Applications:Monitoring protein migration during SDS-polyacrylamide gel electrophoresisMonitoring protein transfer onto membranes after Western blottingSm1841The Thermo Scientific Spectra Multicolor Broad RangeProtein Ladder is a 4-color protein standard containing 10prestained recombinant prokaryotic proteins (10 to 260kDa)for use as size standards in gel electrophoresis and Westernblotting.This prestained protein MW marker is designed formonitoring the progress of SDS-polyacrylamide gelelectrophoresis, for assessing transfer efficiency onto PVDF,nylon and nitrocellulose membranes, and for estimating theapproximate size of separated proteins that have been made visible with gel stains or Western blot detection reagents. Four different chromophores (blue, orange, green, pink) are bound to the different component proteins, producing a brightly colored ladder with an easy-to-remember pattern.Highlights:Size range– 10 proteins spanning 10 to 260kDaMulticolor– four different colors for unambiguous band-size assignmentReady-to-use– supplied in a loading buffer for direct loading on gels; no need to boilSharp bands– color-coded bands of similar intensity for easy visualizationQuality tested – each lot evaluated by SDS-PAGE and Western blottingMembrane-compatible– colored bands transfer to membranes for Western blotting Includes:Dye-stained proteins in 62.5mM Tris-H3PO4 (pH 7.5 at 25°C), 1mM EDTA, 2% SDS, 10mM DTT, 1mM NaN3 and 33% glycerol.Applications:Monitoring protein migration during SDS-polyacrylamide gel electrophoresisMonitoring protein transfer onto membranes after Western blottingSizing of proteins on SDS-polyacrylamide gels and Western blotsProduct Details:Sm1811The Thermo Scientific PageRuler Plus Prestained ProteinLadder is a mixture of nine (9) blue-, orange- andgreen-stained recombinant proteins (10 to 250kDa) for useas size standards in protein electrophoresis (SDS-PAGE)and Western blotting.This prestained protein MW marker is designed formonitoring the progress of SDS-polyacrylamide gelelectrophoresis, for assessing transfer efficiency onto PVDF,nylon and nitrocellulose membranes, and for estimating theapproximate size of separated proteins that have been made visible with gel stains or Western blot detection reagents. A blue chromophore is bound to all proteins, except proteins of two reference bands of 70kDa and 25kDa that are colored with an orange dye and one green reference band of 10kDa. PageRuler Plus Prestained Protein Ladder is ready to use: no heating, further dilution or addition of a reducing agent is required before loading onto a gel.Highlights:Size range– nine proteins spanning 10 to 250kDaReady-to-use– supplied in a loading buffer for direct loading on gels; no need to boilSharp bands– color-coded bands of similar intesity for easy visualizationQuality tested – each lot evaluated by SDS-PAGE and Western blottingBright reference bands– orange at 70 and 25kDa, and green at 10kDaMembrane-compatible– colored bands transfer to membranes for Western blotting Includes:Dye-stained proteins in 62.5mM Tris-H3PO4 (pH 7.5 at 25°C), 1mM EDTA, 2% SDS, 10mM DTT, 1mM NaN3 and 33% glycerol.Applications:Monitoring protein migration during SDS-polyacrylamide gel electrophoresisMonitoring protein transfer onto membranes after Western blottingSizing of proteins on SDS-polyacrylamide gels and Western blotsProduct Details:。
λ-EcoT14Ⅰ说明书
λ-Eco T14Ⅰdigest DNA Marker
使 用 说 明 书
TaKaRa Code ：D3401A
●包 装 量：50μg（500μl×2支；内含1×Loading Buffer）
●浓 度：50 ng/μl
●制品说明：λ-Eco T14 I digest DNA Marker是由Bacteriophage λc I857
Sam7 DNA用Eco T14 I酶切反应后配制而成的。
本制品浓度为50
ng/μl，内含1×Loading Buffer，可根据实验需要，每次取5 μl～
20 μl电泳，使用方便，电泳图像清晰。
●保存温度：-20℃
●使用注意： 1． λDNA digest DNA Markers的原始末端之间经常由COS末
端结合在一起，在电泳前进行热处理（60℃, 5分钟），能使
Marker的电泳图像变得更为清晰。
以λ-Eco T14 I digest DNA Marker为例，原始末端之间的
COS末端的结合，影响19,329 bp和4,254 bp的DNA条带。
如果电泳以后的图像中的4,254 bp的DNA条带亮度低于比其
小的DNA片段时，那说明COS末端的结合情况比较严重。
2．电泳时的加样孔宽度小于6 mm时，每次取5μl制品电泳便可
得到清晰条带。
如果加样孔增宽，须适当增加Marker制品的加
样量。
3． 对DNA电泳而言，Agarose的纯度对DNA条带的清晰度影响
很大。
因此，电泳时应尽量选用质量好的Agarose。
Agarose电泳图像示意图
V2010.04。
TaKaRa 蛋白质分子量标准(低)
∙o Protocolso Standard Protocolo Manual/Datasheet∙Cat.# Product Size Note3450 Protein Molecular Weight Marker (Low) 200 lanes3451 Protein Molecular Weight Marker (High) 200 lanes3452 Protein Molecular Weight Marker (Broad) 200 lanesDescriptionProtein Molecular Weight Markers are designed for use in SDS-Polyacrylamide gel electrophoresis. There are 3 kinds of Protein Molecular Weight Marker, Low (Molecular weight range : 14.3 - 97.2 kDa) , High (Molecular weight range : 44.3 - 200 kDa) and Broad (Molecular weight range : 6.5 - 200 kDa).Each protein is proportioned to yield uniform band intensities when perform staining Coomassie Brilliant Blue R-250 after run on SDS polyacrylamide gel.5µl of 20-fold diluted marker is loaded per lane of SDS-PAGE minigel for standard use. In this case, this marker is sufficient for 200 lanes.Electrophoresis ResultLow(Cat.# 3450) High(Cat.# 3451) Broad(Cat.# 3452)15% SDS-PAGE 7.5% SDS-PAGE 5-20% gradientSDS-PAGEStorageProtein Molecular Weight Marker, 1 MDTT: -20°C5×Loading Buffer : Store at room temperature after used.ComponentProtein Molecular Weight Marker 50 µl5×Loading Buffer1 ml1 M DTT 100 µlConcentration12 µg/µl (Cat.# 3450) 10 µg/µl (Cat.# 3451) 18 µg/µl (Cat.# 3452)Form50 mM Tris-HCl, pH6.81 mM EDTA200mMNaCl50% glycerolComponent Protein Low(Cat.# 3450)Protein Source M.W.(Da)Phosphorylase B Rabbitmuscle97,200Serum Albumin Bovine 66,409Ovalbumin Hen eggwhite44,287Carbonic Bovine 29,000anhydraseTrypsin Inhibitor Soybean 20,100Lysozyme Hen eggwhite14,300High(Cat.# 3451)Protein Source M.W.(Da)Myosin Pig 200,000 β-galactosidaseE. coli116,000Phosphorylase B Rabbitmuscle97,200SerumAlbuminBovine 66,409Ovalbumin Hen eggwhite44,287Broad(Cat.# 3452) < /TR>Protein Source M.W.(Da)Myosin Pig 200,000 β-galactosidase E. coli116,000 Phosphorylase B Rabbit muscle 97,200 Serum Albumin Bovine 66,409Ovalbumin Hen eggwhite44,287CarbonicanhydraseBovine 29,000 Trypsin Inhibitor Soybean 20,100Lysozyme Hen eggwhite14,300Aprotinin Bovinepancreas6,5005×Loading Buffer200 mM Tris-HCl , pH6.810% SDS 0.05%BPB50% GlycerolNote∙All products are intended to be used for research purpose only. They are not to be used for drug or diagnostic purposes, nor are they intended for human use. They shall not to be used products as food, cosmetics, or utensils, etc∙Takara products may not be resold or transferred, modified for resale or transfer, or used to manufacture commercial products without written approval from TAKARA BIO INC.∙If you require licenses for other use, please call at +81 77 543 7247 or contact from our website at ∙Please confirm the content about the license or patent used in this document that relates to the Takara Bio product by clicking the license mark.Moreover, please confirm the "Limited Use Label License" or "patent" concerning the product of another manufacturers or respective owners in their Web site/catalog etc.Page TOP。
Takara说明书
Code No. RR047A 研究用PrimeScript TM RT reagent Kitwith gDNA Eraser(Perfect Real Time)说明书目录内容页码●制品说明1●制品内容 1 ●试剂盒外必备材料 1 ●保存 1 ●特长 2 ●使用注意 2 ●操作方法 2 ●Real Time PCR 4 ●实验例 6 ●附录7 ●关联产品8●制品说明为了准确地进行基因表达量分析，必须满足只有cDNA作为模板检出的先决条件，但Total RNA中常常混有基因组DNA，并可以直接作为PCR反应的模板进行扩增，因此会造成解析结果不准确。
为了避免这种情况发生，通常将检测用引物设计在内含子前后的外显子上，使基因组DNA得不到扩增。
但是，此方法不适合具有单个外显子的基因或两个外显子之间所跨的内含子过小的基因，同时当基因组上有伪基因存在时、或设计引物对基因组有非特异性扩增时、以及基因信息没被完全解析的生物种等也同样不适合于本方法。
在这种情况下，我们常常需要对Total RNA样品进行DNase I处理，以除去残存的基因组DNA。
而DNase I处理通常要进行复杂的纯化操作，同时会造成RNA的降解和损失。
PrimeScript RT reagent Kit with gDNA Eraser是可以除去基因组DNA进行Real Time RT-PCR反应的专用反转录试剂。
Kit中使用了具有较强DNA分解活性的gDNA Eraser，通过42℃，2 min即可除去基因组DNA。
同时由于反转录试剂中含有抑制DNA分解酶活性的组分，经过gDNA Eraser处理后的样品可以直接进行15 min的反转录反应合成cDNA，因此，20 min内即可迅速完成从基因组DNA去除到cDNA 合成的全过程。
使用本制品合成的cDNA适用于SYBR® Green分析法和TaqMan®探针分析法，可以根据实验目的，选择与SYBR®Premix Ex Taq II（Tli RNaseH Plus）、Premix Ex Taq（Probe qPCR）等定量试剂组合使用。
Takara基因组抽提试剂盒说明书
3. 准备 65℃水浴。 4. Solution A、Solution B 若出现沉淀，请于 65℃加热溶解，待恢复至室温后使用。 5. Rinse B 在首次使用前，请添加 56 ml 的 100%乙醇。 6. 洗脱结合于 DNA 制备膜上的基因组 DNA 时，把 Elution Buffer 或灭菌蒸馏水加热至 65℃使用将
●制品内容（50 次量）
本试剂盒分试剂 Set 与 Column Set 两部分。 ■ 试剂 Set
RNase A1（50 mg/ml）*1
20 μl
Lysozyme
60 mg
Glycerol
1.2 ml
EDTA Buffer
2 ml
SP Buffer
10 ml
Solution A*2
12 ml
Solution B*2
注）注意不要残留细小菌块，可以使用振荡器（Vortex） 等剧烈振荡使菌体充分悬浮。
4. 加入 20 μl 的 Lysozyme 溶液，均匀混合后室温静置 5 分钟。
5. 加入 30 μl 的 EDTA Buffer，均匀混合后室温静置 5 分钟。
6. 加入 200 μl 的 Solution A，剧烈振荡后 65℃保温 10
后应充分混合。 ② Lysozyme 失活。Lysozyme 配制成溶液后应保证在-20℃保存。 ③ 洗脱时将灭菌蒸馏水或 Elution Buffer 加热至 65℃后使用将有利于提高洗脱效率。 ④ 严格按照操作方法进行操作。
pEGFP_N1_BMP_2真核表达质粒的构建与鉴定
基金项目:高等学校博士点专项科研基金(编号:20060183050)3通讯作者文章编号:1007-4287(2009)12-1683-03pEGFP 2N1/BMP 22真核表达质粒的构建与鉴定张明磊,常 非,高忠礼,柳 扬,刘光耀3(吉林大学中日联谊医院骨科,吉林长春130033)摘要:目的 构建pEG FP 2N1/BMP 22真核表达质粒,并进行鉴定。
方法 利用RT 2PCR 方法从组织中提取骨形态发生蛋白(BMP 22)的基因片段,与pM D182T 载体连接,构建pM D182T/BMP 22重组质粒,酶切后与pEG FP 2N1真核表达载体连接,构建pEG FP 2N1/BMP 22真核表达质粒,进行测序、酶切鉴定,表明质粒构建成功。
结果 通过测序、酶切鉴定,证明pM D182T/BMP 22质粒构建成功。
结论 本实验成功构建了pEG FP 2N1/BMP 22真核表达质粒,为进一步研究利用BMP 22基因修饰骨组织工程骨种子细胞提供实验基础。
关键词:pEG FP 2N1;骨形成蛋白2;基因中图分类号:R34文献标识码:AConstruction and identification of euk aryotic expression vector pEGFP 2N 1/BMP 22 ZH ANG Ming 2lei ,CH ANG fei ,G AO Zhong 2li ,et al.(China 2Japan Union Hospital o f Jilin Univer sity ,Changchun 130033,China )Abstract :Objective T o construct a eukary otic expression vector carrying BMP 22gene.Methods BMP 22gene was cloned from tissue by using RT 2PCR method and was inserted into pM D182T to constrcuct pM D182T/BMP 22plasmid ,and to construct pEG FP 2N1/BMP 22plasmid by using restriction enzyme.T o identify the plasmid by sequencing and restriction enzyme.R esults The eukary otic expression vector pEG FP 2N1/BMP 22was constructed and identified.Conclusion T o construct the expression vector suc 2cess fully ,BMP 22gene was able to was for bone engineering.K ey w ords :pEG FP 2N1;BMP 22;gene(Chin J Lab Diagn ,2009,13:1683) 在新生骨组织的形成过程中,多种相关细胞因子参与其中并起到十分重要的作用,通过细胞因子的浓度发生改变,继而通过募集祖细胞,发挥调节细胞的增殖和分化等作用。
蛋白Marker说明书
SM0671PageRuler Prestained Protein LadderA prestained SDS-PAGE MW marker with contrasting colored reference bands at10 and 70kDa.The Thermo Scientific PageRuler Prestained Proteis a mixture of ten (10) blue-, orange- and green-srecombinant proteins (10 to 170kDa) for use as sizstandards in protein electrophoresis (SDS-PAGE)Western blotting.This prestained protein MW marker is designed fomonitoring the progress of SDS-polyacrylamide geelectrophoresis, for assessing transfer efficiency onylon and nitrocellulose membranes, and for estimapproximate size of separated proteins that have b visible with gel stains or Western blot detection reagents. The ladder contains one orange refer at 70kDa and one green band at 10kDa.Highlights:•Size range– 10 proteins spanning 10 to 170kDa•Ready-to-use– supplied in a loading buffer for direct loading on gels; no need to boil •Sharp bands– color-coded bands of similar intensity for easy visualization•Quality tested – each lot evaluated by SDS-PAGE and Western blotting•Two reference bands– orange at 70kDa and green at 10kDa•Membrane-compatible– colored bands transfer to membranes for Western blottingIncludes:•Dye-stained proteins in 62.5mM Tris-H3PO4 (pH 7.5 at 25°C), 1mM EDTA, 2% SDS, 1 1mM NaN3 and 33% glycerol.Applications:•Monitoring protein migration during SDS-polyacrylamide gel electrophoresis•Monitoring protein transfer onto membranes after Western blotting•Sizing of proteins on SDS-polyacrylamide gels and Western blotsProduct Details:SDS-PAGE band profile of the Thermo ScientificPageRuler Prestained Protein Ladder. Images arefrom a 4-20% Tris-glycine gel (SDS-PAGE) andsubsequent transfer to membrane.Sm1841Spectra Multicolor Broad Range Protein LadderA prestained, 4-color, SDS-PAGE MW marker especially for proteins between 10 and 260kDa.The Thermo Scientific Spectra Multicolor Broad RangeProtein Ladder is a 4-color protein standard containing 10prestained recombinant prokaryotic proteins (10 to 260kDa)for use as size standards in gel electrophoresis and Westernblotting.This prestained protein MW marker is designed formonitoring the progress of SDS-polyacrylamide gelelectrophoresis, for assessing transfer efficiency onto PVDF,nylon and nitrocellulose membranes, and for estimating theapproximate size of separated proteins that have been made visible with gel stains or Western blot detection reagents. Four different chromophores (blue, orange, green, pink) are bound to the different component proteins, producing a brightly colored ladder with an easy-to-remember pattern.Highlights:•Size range– 10 proteins spanning 10 to 260kDa•Multicolor– four different colors for unambiguous band-size assignment•Ready-to-use– supplied in a loading buffer for direct loading on gels; no need to boil•Sharp bands– color-coded bands of similar intensity for easy visualization•Quality tested – each lot evaluated by SDS-PAGE and Western blotting•Membrane-compatible– colored bands transfer to membranes for Western blotting Includes:•Dye-stained proteins in 62.5mM Tris-H3PO4 (pH 7.5 at 25°C), 1mM EDTA, 2% SDS, 10mM DTT, 1mM NaN3 and 33% glycerol.Applications:•Monitoring protein migration during SDS-polyacrylamide gel electrophoresis•Monitoring protein transfer onto membranes after Western blotting•Sizing of proteins on SDS-polyacrylamide gels and Western blotsProduct Details:SDS-PAGE band profile of the Thermo ScientificSpectra Multicolor Broad Range Protein Ladder.Images are from a 4-20% Tris-glycine gel(SDS-PAGE) and subsequent transfer to membrane.S m1811PageRuler Plus Prestained Protein LadderA prestained SDS-PAGE MW marker with contrasting colored reference bands at 10, 25 and 70kDa.The Thermo Scientific PageRuler Plus Prestained ProteinLadder is a mixture of nine (9) blue-, orange- andgreen-stained recombinant proteins (10 to 250kDa) for useas size standards in protein electrophoresis (SDS-PAGE)and Western blotting.This prestained protein MW marker is designed formonitoring the progress of SDS-polyacrylamide gelelectrophoresis, for assessing transfer efficiency onto PVDF,nylon and nitrocellulose membranes, and for estimating theapproximate size of separated proteins that have been made visible with gel stains or Western blot detection reagents. A blue chromophore is bound to all proteins, except proteins of two reference bands of 70kDa and 25kDa that are colored with an orange dye and one green reference band of 10kDa. PageRuler Plus Prestained Protein Ladder is ready to use: no heating, further dilution or addition of a reducing agent is required before loading onto a gel.Highlights:•Size range– nine proteins spanning 10 to 250kDa•Ready-to-use– supplied in a loading buffer for direct loading on gels; no need to boil•Sharp bands– color-coded bands of similar intesity for easy visualization•Quality tested – each lot evaluated by SDS-PAGE and Western blotting•Bright reference bands– orange at 70 and 25kDa, and green at 10kDa•Membrane-compatible– colored bands transfer to membranes for Western blotting Includes:•Dye-stained proteins in 62.5mM Tris-H3PO4 (pH 7.5 at 25°C), 1mM EDTA, 2% SDS, 10mM DTT, 1mM NaN3 and 33% glycerol.Applications:•Monitoring protein migration during SDS-polyacrylamide gel electrophoresis•Monitoring protein transfer onto membranes after Western blotting•Sizing of proteins on SDS-polyacrylamide gels and Western blotsProduct Details:SDS-PAGE band profile of the Thermo Scientific PageRuler Plus Prestained Protein Ladder. Images are from a 4-20% Tris-glycine gel(SDS-PAGE) and subsequent transfer to membrane.。
TaKaRa DNA 合成指示书
5`末端 Biotin 标记
例 ① ② ③ ④ ⑤ ⑥ ⑦ ⑧
〇
5` 5` 5` 5` 5` 5` 5` 5` 5`
A T g C T g g T C A T T g g
※注意 ●请一定指定制品级别。 特殊要求：
●g 请一定用小写。
●碱基数超过 40 个时请转入第 2 行填写（此时第 2 行的 5`请用斜线划去） 。
TaKaRa DNA/RNA 合成
TaKaRa DNA 合成负责人 收 Fax：0411-87619946 TEL：0411-87641683
依赖页数： 总合成数：
页 支 年
月
日
（注意）请填写清楚以下各栏目，如有没有填写清楚之部分，我们将在电话或传 真确认后再进行合成。此时有可能延误合成时间。
Inosine→ I Mixture→ A,g· · ·R A,T,C· · ·H A,C· · ·M g,T,C· · ·B A,T· · ·W g,A,T· · ·D C,g· · ·S C,T· · ·Y g,T· · ·K A,C,g,T· · ·N
FAX 方向
填写碱基时，为避免错误，请按下列规定填写
Base→ A·C·g·T
A,C,g· 地址 TEL FAX
备 邮编
考
制品级别
OPC 纯化 PAGE 纯化 HPLC 纯化
名称
Base
碱基排列顺序
备考 （标记
3
14
6
9
12
15
18
21
24
27
30
33
36
39
种类、 合成 量指定等）
特巧PCR放大剂盒说明书
･ Unit definition: One unit is the amount of enzyme that will incorporate 10 nmol of dNTP into acid-insoluble products in 30 minutes at 74℃, pH 9.3, with activated soluble salmon sperm DNA as the template-primer.
･ Control Primers 1 and 2 will produce a 6,012 bp amplified DNA fragment from the Control Template (λDNA).
･ Control Primers 1 and 3 will produce a 500 bp amplified DNA fragment from the Control Template (λDNA).
*5 6X Loading Buffer [Composition] 36% 30 mM 0.05% 0.035%
Glycerol EDTA Bromophenol Blue Xylene Cyanol
III. Strials Required but not Provided
･ Reaction mixture for unit definition: 25 mM TAPS (pH 9.3 at 25℃) 50 mM KCl 2 mM MgCl2 0.1 mM DTT
200 μM each dATP･dGTP･dCTP 100 μM [3H]-dTTP 0.25 mg/ml activated salmon sperm DNA
- Agarose gel PrimeGel™ Agarose PCR-Sieve (Cat. #5810A) Agarose L03 [TAKARA] (Cat. #5003)