Clonejet pcr cloning kit pdf
A Basic Polymerase Chain Reaction Protocol . Introduction . The polymerase chain reaction (PCR) is the cardinal laboratory technology of molecular biology. Arguably one of the most powerful laboratory techniques ever discovered, PCR combines the unique attributes of being very sensitive and specific with a great degree of flexibility. Intact Genomics offers high quality competent cells, enzymes, cloning kits, RT-PCR kits, and RT-qPCR kits at very competitive prices. ig™ competent cells have the highest transformation efficiencies in the market. ig™ Taq DNA Polymerase, T4 DNA Ligase, Pfu DNA Polymerase, RT-PCR Kit, SYBR Green qPCR 2x Master Mix, and igScript™ First Strand cDNA synthesis kit have been tested and proven ... GFX PCR DNA and Gel Band Purification Kit is designed for the rapid purification and concentration of PCR products or DNA fragments ranging in size from 50 bp to 10 kb. This kit can be used to purify DNA from reaction volumes up to 100 μl or agarose gel slices up to 900 mg.
genomic DNA of Gladiolus by PCR using the primers GubiP1-1S(5 -ATCGGGCTAAACATAATGGGTGTTCT G-3 ) and GubiP1-1 AS (CTGCGACAGAAAATGTATGG ATCAGTA-3 ). The PCR product was ligated into pGEM-T using AT cloning, transformed in E. coli DH5α, and then sequenced. The clone that contained the ubiquitin gene was named pG1-1. PCR-Ž"?— (3) In-Fusionãï . In-Fusion8 HD Cloning Kit In-Fusion© HD Cloning Kit w/Cloning Enhancer [email protected] HD Cloning Kit w/NucleoSpin [email protected] HD Cloning Kit 300/00FF ¥22,000 ¥101 ,ooo ¥180,000 ¥24,000 ¥195,000 ¥21 ,ooo ¥98,000 ¥1 68,000 ¥70,700 ¥126,000 ¥16,800 ¥79,800 ¥136,500 ¥14,700 ¥68,600 ¥1 17,600 I TA Cloning® Kit Catalog Numbers K2000-01, K2000-40, K2020-20, K2020-40, K2030-01 K2030-40, K2040-01, and K2040-40 . Publication Number MAN0000008 Revision 4.00 . For research use only. Not for use in diagnostic procedures. ... The TA Cloning ® Kit with pCR ... Kit Contents QIAGEN PCR Cloning Kit QIAGEN PCR Cloning Kit (10) (40) Catalog No. 231122 231124 Ligation Master Mix, 2x 50 µl 200 µl pDrive Cloning Vector 0.5 µg 2.0 µg Distilled water 1.7 ml 1.7 ml QIAGEN PCR Cloningplus Kit QIAGEN PCR Cloningplus Kit (10) (40) Catalog No. 231222 231224 BOX I Ligation Master Mix, 2x 50 µl 200 µl The pGEM®-T Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments.
NZY-blunt PCR cloning kit Catalogue number: MB121 01, 24 ligations w / competent cells MB1210 2, 24 ligations Description NZY-blunt PCR cloning kit was designed to allow the direct cloning of PCR products with blunt ends, which result from PCR amplifications … The amplified PCR products were analyzed by electrophoresis on 1% agarose gel in 1X TAE electrophoresis buffer. The amplified products were cloned according to Sambrook and Russell (2001). PCR products were purified from agarose gel using DNA extraction kit (Fermentas) and ligated into pBT, then were transformed into E.coli DH5α. Plasmid ...
The KAPA3G Plant PCR Kit contains KAPA3G DNA Polymerase, a novel, third-generation (3G) enzyme which was engineered via a process of directed evolution for improved tolerance to common plant-derived PCR inhibitors such as polyphenolics and polysaccharides.The KAPA3G Plant PCR Kits are optimized for fast and efficient amplification of plant DNA ... Reaction Cleanup Kit (or similar) (Catalog #28204), using 1 column per pooled sample: Add 4X sample volume of Qiagen Binding Buffer PB to combined PCR. Add 1/100th volume 3M NaOAc. Combined PCR= ~600 uL, Add 2.4 mL PB and 30 uL NaOAc Once PCR is bound to column, Wash 2X w/ 750 uL of PE Perform one dry spin at max speed for 2 minutes The purified PCR product was ligated with the InsT/A vector to generate a recombinant plasmid. Plasmid DNA was purif ied for sequencing using the qiagen QIA prep miniprep kit (Qiagen, United states). Nucleotide sequences were analysed using BLAST analysis and ‘gene tool’ softwares. Fig.1: Schematic representation of cloning of napin promoter
Essential tools for your molecular biology workflows. Our Thermo Scientific molecular biology products are designed to deliver reproducibility and performance in everyday applications including reverse transcription, PCR, nucleic acid electrophoresis, restriction enzyme digests, and cloning. PCR Biosystems, the UK-based PCR experts, have expanded their range of specialist molecular biology kits with the introduction of NGSBIO Library Quant Kit for Illumina®. The new kit contains all the components necessary for accurate and sensitive quantification of DNA libraries prior to next generation sequencing (NGS) with Illumina platforms. TOPO® XL PCR Cloning Reagents TOPO® XL PCR Cloning reagents (Box 1) are listed below. Note that the user must supply the thermostable polymerase. Store Box 1 at -20°C. Item Composition Amount for 10 Reaction Kit Amount for 20 Reaction Kit pCR ®-XL-TOPO 10 ng/µl plasmid DNA in: 50% glycerol 50 mM Tris-HCl, pH 7.4 (at 25°C) 1 mM EDTA 2 mM DTT → JBS dNTP-Mutagenesis Kit #PP-101. Random Mutagenesis by Error-Prone PCR Developed by Caldwell & Joyce (1992) this method introduces mutations in the gene of interest using a PCR reaction under conditions that induce an increased error-rate of the DNA-polymerase. The rate of mutagenesis achieved by error-prone PCR is in the range of 0.6-2.0%. For in vitro use only!. Shipping: shipped at ambient temperature Storage Conditions: store at 4 °C Shelf Life: 12 months Form: Liquid Concentration: 6x conc. Applications: Add one part of Gel Loading Buffer to 5 parts of the DNA sample as shown in the table below. CloneJET™ PCR Cloning Kit #K1231, #K1232 CERTIFICATE OF ANALYSIS #___ Lot __ Expiry Date __ QUALITY CONTROL All components of the kit were function tested in control experiment as described in the manual. A 2 µl aliquot of the ligation mixture was used … The RNAi Core Version 3 (11/12/27) Protocol for shRNA construction-I: PCR method Preparation of cloning vector: 1. Incubate 3 µg of shRNA cloning vector with 5 units (NEB) of EcoRI and 10 units of AgeI, (double digestion) in a reaction volume of 100 µl at 370C for overnight (using NEB #4 buffer). 2.
TOPO TA Cloning® Kit for Sequencing A Limited Use Label License covers this product (see Purchaser Notification). Five-minute cloning of Taq polymerase-amplified PCR products for sequencing Catalog nos. K4530-20, K4575-J10, K4575-01, K4575-40, K4580-01, K4580-40, K4595-01, K4595-40 Version N 19 July 2004 25-0276 1. RT-PCR refers to PCR that uses product of an Reverse Transcription (RT) reaction as template 2. A variant of polymerase chain reaction (PCR) 3. A technique commonly used in molecular biology to detect RNA expression 4. RT-PCR is often confused with real-time polymerase chain reaction (qPCR) 5. RT-PCR is used to qualitatively detect gene ... BigEasy ® Long PCR Cloning Kits Lucigen ® Corporation 6 (888) 575-9695 (608) 831-9011 MA048 v1.7 www.lucigen.com Kit Description The BigEasy Long PCR Cloning Kits contain everything needed to efficiently clone PCR products up to 30 kb into an unbiased, high-fidelity cloning vector. There are two different versions of the kits that are compatible Set preferences, including the number of fragments and the PCR enzyme. Change settings at any time and the results will be instantly updated. ... NEBuilder HiFi DNA Assembly Cloning Kit / Master Mix Manual (PDF) Gibson Assembly Cloning Kit Manual (PDF) ... The NEB PCR Cloning Kit contains an optimized 2X Cloning Master Mix with a proprietary ligation enhancer and a linearized vector that uses a novel mechanism for background colony suppression to give a low background.. It allows simple and quick cloning of any PCR amplicon, whether the amplification reactions are performed with proofreading DNA polymerases, such as Q5® or Phusion® which ... The CloneJET PCR Cloning Kit contains a novel, ready-to-use positive selection cloning vector pJET1.2pJET1.2 sequence/blunt. The vector contains a lethal restriction enzyme gene that is disrupted by ligation of a DNA insert into the cloning site. As a result, only bacterial cells with recombinant plasmids are able to form colonies. Test the success of the PCR by gel electrophoresis and purify the reaction using a PCR cleanup kit, and elute with buffer or water without EDTA. Quantify the concentration of F1 and F2 purifications. Combine F1 and F2 PCR products to a final concentration of 2.8ng/µL of each fragment. Bioneer E-commerce. [Daejeon, Republic of Korea – April 29, 2020] Bioneer specializing in molecular diagnostics announced that it will supply $5.7 million worth of COVID-19 related products to Saudi Arabia.
Shop a large selection of products and learn more about Thermo Scientific™ CloneJET™ PCR Cloning Kit with DH10B Competent Cells Quantity: 40 reactions products 5 CLONE (see page 18) CLONING KITS Perfectly Blunt® Cloning Kits – Efficient “universal” cloning of DNA amplified by any polymerase AccepTor™ Vector Kits – Rapid,direct cloning of DNA amplified with non-proofreading DNA polymerases ANALYZE (see page 22) DNA LADDERS AND MARKERS Perfect DNA™ Ladders and Markers PCR Markers PURIFY (see page 14) DNA PURIFICATION • The CloneJET PCR Cloning Kit is compatible with all PCR buffers supplied by Thermo Scientific. • Gel-analyze the PCR product for specificity and yield before cloning. • Specific PCR products of <1 kb appearing as one discrete band on the gel can be used for ligation directly from PCR reaction mixture without any purification. 10 min. An aliquot of the RT-PCR was resolved by gel electrophoresis (1% agarose) and the appro-priately sized fragments cut from the gel. The DNA was puriﬁed (Geneclean Kit; Bio 101, La Jolla, CA, USA), subcloned into the vector pCRII-TOPO (TA Cloning kit; Invitrogen, San Diego, CA, USA) and transformed into One ShotTM competent cells. The CloneJET™ PCR Cloning Kit is an advanced positive selection system for the highest efficiency cloning of PCR products generated with Pfu DNA polymerase, Taq DNA polymerase or other thermostable DNA polymerases. Additionally, any other DNA fragments, either blunt or sticky-end, can be successfully cloned using the kit. Design Parameters You can design PCR primers from the whole template (= target sequence) or limit the choices to a particular region. For PCR primer pairs, you can specify any required bases at the 3' end of the primer (3' clamp), and a maximum difference in primer melting temperatures. Cholesterol and HDL/LDL Quantification Kit Molecular Enzymes and PCR cloning Kits Growth Factors and Cytokines Polyclonal and Monoclonal Antibodies AOF certified Recombinant Proteins Recombinant Protein A/G and Sepharose Conjugates FOR RESEARCH USE ONLY! Not to be used on humans. Fig. 1. Jurkat cells were cultured in 96-well plate in 100 µl of ...
Amplify PCR product Clone attB-PCR product into pDONR™201 with BP CLONASE™ Enzyme Mix PCR Cloning System (BP CLONASE Mix included) Cat. No. 11821-014 Analyze / Propagate Entry Clone Choose Entry Vector Individual Entry Vectors Cat. Nos. 11813-011; 11816-014; 11817-012; 11818-010; 11819-018 Ligation of restriction fragment into Entry Vector The cloning of PCR products into plasmids is a routine step in many molecular biology processes. The DynaExpress TA PCR Cloning Kit is a PCR cloning kit based on T-A pairing of PCR products and T vector. This kit is manufactured by new technologies of ligation and transformation. The 2 × Ligation Buffer Description: Description The GenScript CloneEZ ® PCR Cloning Kit is designed for the quick PCR cloning , it is especially powerful in high-throughput cloning of PCR products into any destination vector effectively by sidestepping the tedious and limiting tasks such as selecting proper restriction enzymes, phosphatases, or ligases. Using our proprietary CloneEZ ® Enzyme, this kit is able to ...