Previous Articles Next Articles A Highly Efficient Protocol for Transformation of Saccharomyces cerevisiae and Pichia pastoris by Electroporation Click on each item for the direct links to … I had some trouble transforming Pichia, and finally managed some good transformants by increasing amount of linealized DNA used in electroporation, Pichia is tricky to transform, you should be using 1-5 ug DNA aprox. 6. How to sterilize disposable electroporation cuvettes - For Pichia Pastoris transformation (Jun/28/2005 ) Hi,all: Recently I was trying to use an electroporation system to transform Pichia Pastoris, Since some of my colleagues here use 75% alcohol to sterilize the disposable electroporation … For control electroporation dilute pUC19 to 10 pg/µl with Milli-Q water. We report here that pretreatment of P. pastoris with 0.1 M lithium acetate (LiAc) and 10 mM dithiothreitol (DTT) before electroporation increased transformation efficiency approximately 150-fold. The instrument features an intuitive operation and user-friendly programming of standard methods. 4308 915.545 - Pichia pastoris. VWR #60818-667) at room temperature. The yeast Pichia pastoris is widely used for expression of foreign proteins. Turn on electroporator and set to 1.7-2.5 kv (optimize for strain), 200 ohms and 25 µF. 3.4.2 Electroporation using a Pre-Set Protocol.....22 3.4.3 Modifying Pre-Set Protocol Parameters ........................................25 3.4.4 Saving Changes to Pre-Set Protocols............................................25 MaxCyte® STX electroporation transfection apparatus. This book focuses on recent developments of Pichia pastoris as a recombinant protein production system. A: Yes. We reasoned that there were no reason why transformants could not be selected in liquid rather than in solid phase (i.e., on agar) provided the selection reagent Zeocin was included in the culture medium. Gram-positive bacteria such as Streptococcus pneumoniae and Lactobacillus No. Electroporation of gram-negative bacterial strains can achieve transformation success rates in the range of 1x1010 transformants/μg DNA. It also can be used as a reference method for other Bacillus strains that are refractory to 4. Electroporation device and 0.2 cm cuvettes or reagents for transformation 16°C, 37°C, and 65°C water baths or temperature blocks 30°C and 37°C shaking and non-shaking incubators Transformation efficiencies for Pichia pastoris are usually several orders of magnitude below those for other yeast. Yeasts, such as Saccharomyces cerevisiae, Schizosaccharomyces pombe and Pichia pastoris, are routinely used in biology research labs around the world. The Pichia EasyComp™ Transformation Kit is a unique kit designed for the rapid preparation and transformation of competent Pichia pastoris cells. The Pichia EasyComp™ Transformation Kit offers several advantages over commonly used transformation procedures such as spheroplast formation: Thermo Fisher easyselect pichia expression kit manual Easyselect Pichia Expression Kit Manual, supplied by Thermo Fisher, used in various techniques. Electroporation is widely used for highly efficient transformation of ... 2010, Vol. This method is well suited for the cloning and expression of huge DNA fragments such as gene clusters in B. thuringiensis. Pichia Protocols focuses on recent developments of Pichia pastoris as a recombinant protein production system. Electroporation Protocol. I had some trouble transforming Pichia, and finally managed some good transformants by increasing amount of linealized DNA used in electroporation, Pichia is tricky to transform, you should be using 1-5 ug DNA aprox. The electroporation protocol will vary depending on the strain so this protocol may need to be optimized. Transformation efficiencies for Pichia pastoris are usually several orders of magnitude below those for other yeast. 2 Methods Cloning into pPIC6 A, B, and C General Molecular Biology Techniques For assistance with E. coli transformations, restriction enzyme analysis, DNA biochemistry, and plasmid preparation, refer to Molecular Cloning: A Laboratory Manual (Sambrook et al., 1989) or Current Protocols in Molecular Biology (Ausubel et al., 1994). Production of enzymes in Pichia pastoris. A resource portal for protocols in bioscience including molecular biology, cell biology, immunology, plant biology, developmental biology, neuroscience, bioinformatics, genetics However the most popular electroporation protocol for Pichia pastoris is the one by Lin-Cereghino et al. Before plating out aliquots on corresponding selective media, the cells (80 µl) were allowed to regenerate for two hours at 28°C with 500 µl 1 M sorbitol and 500 µl YPD. Select the appropriate Expression Vector for your target gene. In Pichia Protocols 2 nd Edition, the author focuses on new information that has come to light in recent years but repeats key basic methods so that researchers are not absolutely dependent on having both editions in order to utilize the system. 3. Q: Is the expression of a recombinant protein in shake flask directly transferable to a fermentor? Section 11 Electroporation of Pichia pastoris ... Electroporation of E. coliis generally carried out at a voltage of 1.8 kV (E = 18 kV/cm) when electroporating cells in 0.1 cm cuvettes and at a voltage of 2.5 kV (E = 12.5 kV/cm) when electroporating cells in 0.2 cm cuvettes. Abstract: The Chemicals, Equipments & Supplies box on the right contains a list of materials used in this protocol. Transformation efficiencies for Pichia pastoris are usually several orders of magnitude below those for other yeast. This article provides detailed basic protocols for the expression of heterologous genes and the synthesis of recombinant proteins utilizing this yeast. If you do not have access to an electroporation device, you may use the Pichia Spheroplast Kit (Cat. The recombinant expression plasmid, designated as pPIC9K-hly, was linearized with SacⅠ and transformed into Pichia pastoris GS115 (his4, Mut+) by electroporation. Not for use in diagnostic procedures. Pichia pastoris has … 49 ›› Issue (3): 98-101. Medical books Pichia Protocols . Y1000 with 1% yeast extract, 2% peptone, 2% glucose and 2% agar) and incubating for 2 days at 30°C. The following protocol can be a reference for you to understand more details of the techniques utilized in our lab and workflow involved in producing the protein of interest using P.pastoris. After completing washing steps, 178 incubate the tube on ice for 3-5 min, mix with DNA (PCR product or digested plasmid) and 179 gently mix the content of tube. Pichia Protocols (Methods in Molecular Biology) This book focuses on recent developments of Pichia pastoris as a recombinant protein production system. Introduction Pichia pastoris is a methylotrophic yeast largely used for heterologous protein production in biotechnology. Condensed protocol for competent cell preparation and transformation of the methylotrophic yeast Pichia pastoris. Cell density and the pichia pastoris electroporation protocol reduce the different purities. The standard way of electroporating Pichia is to follow the Invitrogen protocol, which is copied pretty much verbatim here. 600 of ~3.0. Each chapter presents cutting-edge and cornerstone protocols for utilizing P. pastoris as a model recomibinant protein production system. Highlighted topics include a discussion on the use of fermentors to grow Pichia pastoris, information on the O- and N-linked glycosylation, methods for labeling Pichia pastoris There are many other protocols available, but by far the most important consideration is the fact that you should use fresh Pichia competent cells. 1. For control electroporation dilute pUC19 to 10 pg/µl with Milli-Q water. Strains are in YPD + 1M Sorbitol broth and are shipped at ambient temperature. Receive all current lab protocols on inbox free by entering your email address: ... Electroporation of Pichia pastoris. 7. We recommend you prepare a set of glycerol master stocks prior to using your Pichia (page 9) and TOP10F´ E. coli cells (page 10). coli, Agrobacterium tumerfaciens, Pichia pastoris, and Saccharomyces cerevisiae. Inoculate 250 ml YPD with an aliquot of the overnight culture to reach an O.D. The electroporation protocol will vary depending on the strain so this protocol may need to be optimized. Invitrogen protocols strongly recommend to grow strains at 28ºC, to prevent overwarming in cultured flasks (be aware that even if temperature inside the shaker is 30ºC, the flask content can get warmer due to yeast growth). Feb 08, 2015Here jenny nimmothe snow spider , we present this electroporation protocol, For reference, a normal field strength used for bacteria and yeast electroporation is 1. 5 kVcm. For the other procedures, readers are referred to either of the volumes of Methods in Molecular Biology: Pichia Protocols (Cregg, 2008; Higgins and Cregg, 1998). ZERO BIAS - scores, article reviews, protocol conditions and more Place recovery SOC in 37°C water bath. Highlighted topics include a discussion on the use of fermentors to grow Pichia pastoris, information on the O- and N-linked glycosylation, methods for labeling Pichia pastoris expressed proteins for structural studies, and the introduction of mutations in Pichia pastoris genes by … Pichia Expression Kit For expression of recombinant proteins in Pichia pastoris Catalog Number K1710-01 Revision A.0 Publication Number MAN0000012 . 7. Streak Pichia cells from a glycerol stock (30 % glycerol) onto YPD plates to grow single colonies. Click on each item for the direct links to … No. ii Research Use Only. Highlighted topics include a discussion on the use of fermentors to grow Pichia pastoris, information on the O- and N-linked glycosylation, methods for labeling Pichia pastoris expressed proteins for structural studies, and the introduction of mutations in Pichia pastoris genes by … pichia electroporation device too diluted for pichia do you a link was spun down. Prepare the Sorting Plate 7 Prepare a 96 well culture plate to receive cells following Nucleofection ... Protocol No. Prepare 17 mm x 100 mm round-bottom culture tubes (e.g. Electroporation is the application of controlled electrical pulses to living cells in order to permeabilize the cell membrane for the purposes of transfection or transformation. Low transformation efficiencies have been accepted as inevitable for Pichia vectors because, unlike many other yeast systems, the plasmid must both enter the yeast cell and integrate into specific locations in the host chromosome, an inherently inefficient process (4). Pichia pastoris Protocol for Growth in a Fermentor 2 of 12 occur. As a rule of thumb, using a “standard” fermentation condition, it is possible to increase expression by about 5 to 10 fold from shake flask. Therefore, researchers have recently turned to eukaryotic yeast and mammalian expression systems for protein production. Several μg of MDA products was transformed by electroporation into Pichia cells, and the activities of extracellularly expressed PcCel6A mutants towards crystalline and amorphous celluloses were compared with those of wild-type enzyme to identify key amino acid residues affecting degradation of crystalline cellulose. Based on EasySelect™ Pichia Expression Kit (Invitrogen), to increase the efficacy of external DNA integration into the Pichia genome, first, the cloned vector should be linear with restriction enzymes such as Sac I, PmeI, and BstX I. How to sterilize disposable electroporation cuvettes - For Pichia Pastoris transformation (Jun/28/2005 ) Hi,all: Recently I was trying to use an electroporation system to transform Pichia Pastoris, Since some of my colleagues here use 75% alcohol to sterilize the disposable electroporation … Transformation of Pichia pastoris cells for the introduction of the gene of interest is common practice for expression and purification of a heterologous protein(s). Electroporation of Pichia 2003-09-17 (Based on Invitrogen protocol) Materials • 5-100 ug pure plasmid DNA containing gene of interest, linearized by restriction digestion (p270 can be linearized with SacI, PmeI, and BstX) • Extra YPD media • YPDS plates (with proper antibiotic, ie, 100 and 500 ug/mL Zeocin) pichia pastoris strain gs115 cells were transformed with linearized plasmids by electroporation according to the user manual for the multi- copy pichia expression kit ( invitrogen). Yeasts are easy-to-culture, unicellular eukaryotes, and make excellent model organisms because of the similarity of their genes and proteins with those of their mammalian counterparts. These pulses are delivered to a pair of electrodes by a pulse generator. Pictured is the apparatus set up for 100mL flow electroporation … Using the following protocol, electroporation with 2 ul of the 5 ng/ul control The Pichia EasyComp™ Transformation Kit is a unique kit designed for the rapid preparation and transformation of competent Pichia pastoris cells.The Pichia EasyComp™ Transformation Kit offers several advantages over commonly used transformation procedures such as spheroplast formation: • Competent cells can be frozen for future use, minimizing waste Pichia Pastoris Expression System One such eukaryotic yeast is the methanoltrophic Pichia pastoris. DNA was linearized at a unique restriction site in the AOX1 region. PEI Electroporation . The use of Pichia Pastoris as a recombinant protein system has increased greatly in popularity over the years. Addgene Protocols On this page you'll find practical lab protocols that you can use for a wide range of applications, with videos for select protocols in the right-hand column. SHORT PROTOCOL I No. Pichia pastoris transformation through electroporation protocol (method) by Igem Dusseldorf Wu, S. and Letchworth, G. J. Yeast cells are used to study gene function, … Spread 50, 100, and 200 µl to both YPDS + 250 ug/ml Zeocin and … we are trying to express a protein following invitrogen pichia expression kit manual using extracellular vector ppic9k in pichia pastoris without any tag. Highlighted topics include a discussion on the use of fermentors to grow Pichia pastoris, information on the O- and N-linked glycosylation, methods for labeling Pichia pastoris expressed proteins for structural studies, and the introduction of mutations in Pichia pastoris genes by … Inoculate 10 ml YPD with one colony and grow 12-14 h with shaking at 30 °C to an O.D. orders pichia protocols methods in molecular biology pichia protocols methods in molecular biology pdf epub ebook d0wnl0ad this book focuses on recent developments of pichia pastoris as a recombinant protein production system highlighted topics include a discussion on the use of fermentors to grow pichia pastoris information on the o. Vectors that are designed to integrate into the Pichia chromosome enable stable expression, but transformation efficiencies are very low (1–3). This volume fully updates and expands upon the first edition of Pichia Protocols, and focuses primarily on information that has come to light since its original publication. Abstract. 5. Protocols. We use a small, nonrefrigerated centrifuge capable of spinning six 50-mL conical tubes at a time, enabling us to quickly prepare competent cells of six different strains simultaneously. Figure 2. (2004) High efficiency transformation by electroporation of Pichia pastoris pretreated with lithium acetate and dithiothreitol. As an expression system, the main advantages of P. pastoris over E. coli are that the eukaryotic P. pastoris is capable of car- rying out post-translational modifications, such as glycosyl- Highlighted topics include a discussion on the use of fermentors to grow Pichia pastoris, information on the O- and N-linked glycosylation, methods for labeling Pichia pastoris The electroporation procedure is most commonly used and therefore a modified version of that described by (Becker and Guarante, 1991) will be outlined in detail. 7. Pichia pastoris Protocol for Growth in a Fermentor 2 of 12 occur. Download: Protocol – Protocol No. @article{Wu2004HighET, title={High efficiency transformation by electroporation of Pichia pastoris pretreated with lithium acetate and dithiothreitol. Electroporation Protocol. The electroporation protocol will vary depending on the strain so this protocol may need to be optimized. For control electroporation dilute pUC19 to 10 pg/µl with Milli-Q water. Turn on electroporator and set to 1.7-2.5 kv (optimize for strain), 200 ohms and 25 µF. Transfer the content of tube in pre-labeled electroporation 180 cuvettes which were already kept on ice. tive electroporation protocol for large plasmids and wild type strains of B. thuringiensis. 2. You can also have your target protein secreted into the culture media if your protein is generally secreted, glycosylated or directed to an intrac… Abstract: The Chemicals, Equipments & Supplies box on the right contains a list of materials used in this protocol. K172001), PEG 1000 protocol (page 78 of the manual), LiCl protocol (page 80 of the manual), or the Pichia EasyComp™ Transformation Kit (Cat. Based on EasySelect™ Pichia Expression Kit (Invitrogen), to increase the efficacy of external DNA integration into the Pichia genome, first, the cloned vector should be linear with restriction enzymes such as Sac I, PmeI, and BstX I. US9200270B2 US12/380,703 US38070309A US9200270B2 US 9200270 B2 US9200270 B2 US 9200270B2 US 38070309 A US38070309 A US 38070309A US 9200270 B2 US9200270 B2 US 9200270B2 Authority US United States Prior art keywords yeast dna vector electroporation cells Prior art date 2008-03-03 Legal status (The legal status is an assumption and is not a legal conclusion. Expectations Each “one-shot” tube of BioGrammatics electroporation-competent Pichia pastoris (E-comp) cells has 25 – 30 ul of frozen competent cells. Incubate transformations for 1-2 hours at 30°C and 200 rpm. Also, minimal amount of resuspending … 2) Pellet and resuspend in 10 mL of ice-cold 100mM Tris, 10mM EDTA buffer with 200 DTT. Must both enter the yeast cell Pichia is usually transformed by electroporation or by the timecon Free Plasmid Maxi Kit protocol and reagents. BioTechniques 36 (1), 152 – 154, DOI: 10.2144/04361DD02 Pichia Pastoris Expression System One such eukaryotic yeast is the methanoltrophic Pichia pastoris. Over the last couple of decades, methylotrophic yeast, Pichia pastoris emerges as an important yeast species owing to its increasing application in industry and basic biological research. Cold Spring Harbor Protocols is now offering free full-access trial subscriptions. Immediately add 0.5 ml of ice-cold 1M Sorbitol to the electroporation cuvette, and transfer cells to a round bottom culture tube with YPD broth. Transformants were plated on the appropriate medium (RDB minus histidine or YPDS plus 100 µg/ml Zeocin™ agent). Electroporation Protocol. The introduction of foreign DNA into the yeast Pichia pastoris is similar to the processes used with Saccharomyces cerevisiae and routinely performed by the electroporation of plasmid DNA into electro-competent (E-comp) P. pastoris cells (Becker and … It is important to have healthy log phase cells. Then, linear DNA should be inserted into the competent cells by electroporation. Highlighted topics include a discussion on the use of fermentors to grow Pichia pastoris, information on the O- and N-linked glycosylation, methods for labeling Pichia pastoris expressed proteins for structural studies, and the introduction of mutations in Pichia pastoris genes by … 5 kVcm. In the case of a Mut – strain, Pichia is unable to metabolize methanol. For the other procedures, readers are referred to either of the volumes of Methods in Molecular Biology: Pichia Protocols (Cregg, 2008; Higgins and Cregg, 1998). Vectors can be maintained as autonomously replicating elements or integrated into the P. Bioz Stars score: 94/100, based on 1 PubMed citations. GS115 P. pasto-ris cells were transformed by electroporation with the linearized constructs. For the other procedures, readers are referred to either of the volumes of Methods in Molecular Biology: Pichia Protocols (Cregg, 2008, Higgins, D. R., and Cregg, J. M. 1998). Invitrogen’s protocol implies plating cells on Zeocin agar plates after electroporation for selecting transformants. Therefore, researchers have recently turned to eukaryotic yeast and mammalian expression systems for protein production. This article provides detailed basic protocols for the expression of heterologous genes and the synthesis of recombinant proteins utilizing this yeast. Aims: To elaborate an effective electroporation protocol for large plasmids and wild type strains of Bacillus thuringiensis. Then, linear DNA should be inserted into the competent cells by electroporation. Pichia Protocols focuses on recent developments of Pichia pastoris as a recombinant protein production system. Place recovery SOC in 37°C water bath. You may choose to express your protein intracellularly if your protein is cytosolic and non-glycosylated. Methods and Results: The effect of DNA desalting, wall-weakening agency, cell growth conditions, electroporation solutions, and electric fields on electroporation efficiency was evaluated to optimize electroporation conditions for B. thuringiensis. This lecture explains about the electroporation technique. It also explains the role of electroporation in transformation. The electroporation proce- dure is most commonly used and therefore a modified version of that described by (Becker and Guarante, 1991) will be outlined in detail. Also, minimal amount of resuspending … When received, strains should be stored at 4°C and revived within 1 week by streaking for single colonies to YPD Agar plates (Teknova, Cat. Feb 08, 2015Here jenny nimmothe snow spider , we present this electroporation protocol, For reference, a normal field strength used for bacteria and yeast electroporation is 1. Approximately 5 μg of the MDA product (50 μl of the MDA product) was digested with BlnI prior to transformation of P. pastoris. No. We report here that pretreatment of P. pastoris with 0.1 M lithium acetate (LiAc) and 10 mM dithiothreitol (DTT) before electroporation increased transformation efficiency approximately 150 … The Eppendorf Eporator is a compact instrument designed for fast and controlled electroporation of bacteria, yeasts and other microorganisms. One of the newest and most promising of these is the yeast Pichia pastoris. (2005). Pichia pastoris has … The cloned cDNA (hly) was 393 bp in length, encoding a 130 amino acid hLY with a calculated molecular mass of 14,698 Da. Turn on electroporator and set to 1.7-2.5 kv (optimize for strain), 200 ohms and 25 µF. Pichia Protocols focuses on recent developments of Pichia pastoris as a recombinant protein production system. Pichia pastoris is a methylotrophic yeast increasingly important in the production of therapeutic proteins. Receive all current lab protocols on inbox free by entering your email address: ... Electroporation of Pichia pastoris. These electroporation Pichia is usually transformed b… One of the newest and most promising of these is the yeast Pichia pastoris. Electroporation is widely used for highly efficient transformation of Saccharomyces cerevisiae.As many strains degenerate over time or over environmental exposure,their electroporation efficiencies could drop markedly by 2 to 3 orders of magnitude.Here we describe a modified electroporation procedure which included single stranded carrier DNA for electroporation as well as a post-electroporation … The electroporation proce- dure is most commonly used and therefore a modified version of that described by (Becker and Guarante, 1991) will be outlined in detail. Apparently protein expression in Pichia is quite sensitive to temperature, and stops above 32ºC. We report here that pretreatment of P. pastoris with 0.1 M lithium acetate (LiAc) and 10 mM dithiothreitol (DTT) before electroporation increased transformation efficiency approximately 150-fold. For Pichia pastoris, the fate of introduced DNAs is generally similar to those described for Saccharomyces cerevisiae. The key to the molecular genetic manipulation of any organism is the ability to introduce and maintain DNA sequences of interest. BTX generally recommends a system that has an exponential decay waveform for bacteria, plant cells, insect cells, and yeast applications, and a square waveform for mammalian cell work. 1. What is Electroporation? Electroporation has become the research standard for the introduction of biological material into cells. 35 I Page 2 1. Furthermore, unlike the electroporation cell preparation procedure, our condensed protocol does not require a large, refrigerated centrifuge. Electroporation of BioGrammatics competent Pichia cells, 2017-06-01. bG-Customer Protocol – 0XX. Prepare Nucleofector System 5 Turn on the nucleofector and load the X-core and the program of the desired cell type (or closest cell type) Resuspend the Alt-R Cas9 Electroporation Enhancer 6 Resuspend the Alt-R Cas9 Electroporation Enhancer to 100 uM in IDTE. Typically, the most commonly transformed cell lines are bacteria and yeast, such as Escheria coli, Agrobacterium Tumerfaciens, Pichia pastoris and Saccharomyces cerevisiae. Electroporation of these gram-negative bacterial strains can achieve transformation success rates in the range of 1X10 10 transformants/µg DNA. Figure 1. You can find much more video content on the Addgene Video Page . Must both enter the yeast cell Pichia is usually transformed by electroporation or by the timecon Free Plasmid Maxi Kit protocol and reagents. After purification of DNA by ethanol precipitation, electroporation was carried out according to the instruction manual of the EasySelect™ Pichia expression kit. Pichia pastoris Protocol for Growth in a Fermentor 2 of 12 occur Therefore, researchers have recently turned to eukaryotic yeast and mammalian expression systems for protein production Pichia Pastoris Expression System One such eukaryotic yeast is the methanoltrophic Pichia pastoris Pichia pastoris has … US5879891A US08/932,004 US93200497A US5879891A US 5879891 A US5879891 A US 5879891A US 93200497 A US93200497 A US 93200497A US 5879891 A US5879891 A US 5879891A Authority US United States Prior art keywords transformation lithium acetate dtt dna electroporation Prior art date 1997-09-17 Legal status (The legal status is an assumption and is not a legal conclusion. Protocol 505b Electro Cell Manipulator ™ ECM ® 830 ELECTROPORATION PROTOCOL Cell line: Pichia pastoris Transfectant: plasmid DNA pGAPZ α Cell Preparation: 1) Grow cells to an OD 600 of between 0.5 to 0.8 in a 50-mL culture. K173001). The Pichia and TOP10F´ E. coli stabs supplied with the kit are guaranteed until the expiration date marked on tube when stored at 4°C. The transformations (100 ng circular plasmid DNA/CRISPR-Cas9 plasmids, one μg linear plasmid DNA/HR donor cassettes) were performed using a condensed electroporation protocol (Lin-Cereghino et al., 2005) and grown for two days on YPD (1% w/v yeast extract, 2% w/v peptone and 2% w/v glucose) plates (1.5% agar) containing 100 μg/ml Zeocin. This book focuses on recent developments of Pichia pastoris as a recombinant protein production system. Pichia Protocols focuses on recent developments of Pichia pastoris as a recombinant protein production system. Electroporation Protocol (C2986) Protocols.io also provides an interactive version of this protocol where you can discover and share optimizations with the research community.. Protocol. Experimental protocol scheme comparing PEI and MaxCyte® STX electroporation transient transfection techniques.. 2 Publication 5423-025-REV 1.0 • www.btxonline.com Research Only BTX 84 October Hill Rd Holliston, MA 01746, USA Phone: 1-508-893-8999 Fax: 1-800-429-5732 Web: www.btxonline.com Warranty BTX warranties the ECM 830 Electroporation Systems for a 1. Pichia vector and the Zeocin™-resistant pPICZ B Pichia vector. High efficiency transformation by electroporation of Pichia pastoris pretreated with lithium acetate and dithiothreitol. P. pastoris competent cells were prepared with the condensed protocol and transformations were performed by electroporation, essentially as described by .

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