Warning: file_get_contents(http://94.23.197.202:8002/search): failed to open stream: HTTP request failed! HTTP/1.0 500 (No room for thread in thread queue) in /home/lookformedical/www/web.php on line 237

Warning: file_get_contents(http://94.23.197.202:8002/search): failed to open stream: HTTP request failed! HTTP/1.0 500 (No room for thread in thread queue) in /home/lookformedical/www/web.php on line 237

Warning: file_get_contents(http://94.23.197.202:8002/search): failed to open stream: HTTP request failed! HTTP/1.0 500 (No room for thread in thread queue) in /home/lookformedical/www/web.php on line 237

Warning: file_get_contents(http://94.23.197.202:8002/search): failed to open stream: HTTP request failed! HTTP/1.0 500 (No room for thread in thread queue) in /home/lookformedical/www/web.php on line 237
*  A Modified MultiSite Gateway Cloning Strategy for Consolidation of Genes in Plants | SpringerLink
The genome information is offering opportunities to manipulate genes, polygenic characters and multiple traits in plants. Although a number of approaches have been developed to manipulate traits in plants, technical hurdles make the process difficult. Gene cloning vectors that facilitate the fusion, overexpression or down regulation of genes in plant cells are being used with various degree of success. In this study, we modified gateway MultiSite cloning vectors and developed a hybrid cloning strategy which combines advantages of both traditional cloning and gateway recombination cloning. We developed Gateway entry (pGATE) vectors containing attL sites flanking multiple cloning sites and plant expression vector (pKM12GW) with specific recombination sites carrying different plant and bacterial selection markers. We constructed a plant expression vector carrying a reporter gene (GUS), two Bt cry genes in a ...
  https://link.springer.com/article/10.1007%2Fs12033-012-9499-6
*  Help with cloning - Molecular Cloning - BioForum
Help with cloning - posted in Molecular Cloning: Hi, I want to generate a plasmid for mammalian expression in which my reporter gene is downstream of an E2F1 promoter sequence. The standard mammalian expression vectors have multiple cloning sites downstream of CMV promoters, to ensure that I only see expression of my gene upon E2F1 transactivation do I need to remove the CMV promoter?? Or, does anyone know of any vectors which have no promoter sites that I could use instead?? Thanks! P
  http://www.protocol-online.org/forums/topic/8713-help-with-cloning/
*  TOPO vs. Gateway cloning - Molecular Cloning - BioForum
TOPO vs. Gateway cloning - posted in Molecular Cloning: Ok, I have been at this for a while now, and I think it's time to upgrade my technology so I can get some results. I am attempting to clone a gene of interest from potato. The fragment is about 6.5 Kb, and it includes the coding sequence, and 3Kb upstream (presumably the native promoter.) The overall goal is to get this chunk of DNA into a binary vector and transform plants with it. What I've already tried: -a-tailing and cl...
  http://www.protocol-online.org/forums/topic/21526-topo-vs-gateway-cloning/
*  cloning vector,blunt zero cloning,pEASY®-Blunt Zero Cloning Kit,Cloning Vectors,Beijing TransGe
pEASY®-Blunt Zero Cloning Kit,Cloning Vectors,Cloning and Mutagenesis System,Products,Beijing TransGen Biotech Co.Ltd,OverviewContents& storageCitations & referencesRelated ImagesDownloadOverviewDescriptionpEASY ®-Blunt Zero Cloning Vector c
  http://www.transbionovo.com/rx_detail/productId=208.html
*  Manufacturing Biotherapeutics - Starting with CHO Cloning - Cell Culture Dish
For every Herceptin, for every Enbrel, Humira and Rituxan, and for every successful biotherapeutic product in the market today, there is a long history of its product development. We all hope that our target protein product will make a difference in both the therapeutic market and the lives it will eventually affect as it reaches its ultimate goal of becoming a human therapeutic. The reality is, in order for it to be a feasible therapeutic, the protein must first be manufactured. And one of the very first steps in determining the manufacturing capability of the product is the crucial step of choosing the production cell line that will produce this protein.. There are many challenges in single cell CHO cloning procedures to obtain valuable stable CHO production cell lines. We often rely on efficient cloning strategies and cloning media to execute such a crucial task in the developmental process of a recombinant protein product. An important aspect of ...
  http://cellculturedish.com/2011/03/manufacturing-biotherapeutics-starting-with-cho-cloning/
*  Genomic -Lactamase? Blue White screening not working - Molecular Cloning - BioForum
Genomic -Lactamase? Blue White screening not working - posted in Molecular Cloning: Hi everyone, Im having some trouble regarding my cloning procedure. In the past we used to have some XL-Blue competent E.Coli for transformation. Now we switched to another commercial strain (http://tools.lifetec...h5alpha_man.pdf) For cloning I always subclone my PCR products first into pGEM-T vector, transform my bacteria and use blue-white screening to select positively transformed clones....
  http://www.protocol-online.org/forums/topic/32700-genomic-ss-lactamase-blue-white-screening-not-working/
*  Trouble with TOPO-Cloning - Molecular Cloning - BioForum
Trouble with TOPO-Cloning - posted in Molecular Cloning: Hi, Im trying to perform a Gateway Cloning procedure. I have tried to clone a PCR Product into an entry vector via TOPO-Cloning, but, according to my gel electrophoresis it is not working. Here is an overview of what I have done: Day 1: 3.3kb sequence amplified via PCR Product --| checked with gel, worked. Day 2: Gel was extracted and DNA isolated via Qiagen Gel Extraction Kit --| measured sufficient amount of DNA, pro...
  http://www.protocol-online.org/forums/topic/34623-trouble-with-topo-cloning/
*  Molecular Cloning - BioForum - Page 70
Molecular Cloning: Any techniques related to molecular cloning including restriction digestion, ligation, transformation, plasmid...
  http://www.protocol-online.org/forums/forum/42-molecular-cloning/page-70?prune_day=100&sort_by=Z-A&sort_key=last_post&topicfilter=all
*  Gene Cloning: A breakthrough - Orbit Biotech
During the last few decades, techniques for manipulating eukaryotic as well as prokaryotic DNA have witnessed a remarkable development. There are three main phases of the development of these techniques, which include recombinant DNA technology and gene cloning; polymerase chain reaction and DNA chips and microarrays. Using recombinant DNA technology, we can isolate and clone single copy of a gene or a DNA segment into an indefinite number of copies, all identical. This became possible, because bacteria, phages and plasmids reproduce in their usual style, even after insertion of foreign DNA, so that the inserted DNA also replicates faithfully with the parent DNA. This technique is called gene cloning. It involves the production of a large number of identical DNA molecules from a single ancestral DNA molecule. The essential characteristic of gene cloning is that the desired gene or DNA fragments must be selectively amplified resulting in a large increase in ...
  http://orbitbiotech.com/gene-cloning-a-breakthrough/
*  Molecular Cloning - BioForum
Molecular Cloning: Any techniques related to molecular cloning including restriction digestion, ligation, transformation, plasmid...
  http://www.protocol-online.org/forums/forum/42-molecular-cloning/?prune_day=100&sort_by=Z-A&sort_key=last_post&topicfilter=all
*  cloning trouble - Molecular Cloning - BioForum
Page 1 of 2 - cloning trouble - posted in Molecular Cloning: Hello all, I'm quite new at cloning.. i'm using a kit from invitrogen. the pcr product i want to clone is 5 kb and i'm using chemically competent cells which i've stored in a -80 freezer. My first two trials yielded no results-no colonies- so i began to doubt the competency of the cells (there have been power failures), the second time around i plated some on Macconkey. I got a few colonies and I've inoculate...
  http://www.protocol-online.org/forums/topic/6967-cloning-trouble/
*  Two genes, same MCS - Molecular Cloning - BioForum
Two genes, same MCS - posted in Molecular Cloning: Sorry if this is a repeat topic, I have looked/searched. I have a vector that I am trying to give dual resistance to depending on cell line I want to express in. A coworker said I could just put the puromycin cassette into the MCS but I am just wondering if this will cause problems? Two genes in one multiple cloning site? It's a C1 vector but if I put a stop codon after the first gene, will the second one still express?
  http://www.protocol-online.org/forums/topic/29020-two-genes-same-mcs/
*  plasmid, BAC, or cosmid? - Molecular Cloning - BioForum
plasmid, BAC, or cosmid? - posted in Molecular Cloning: Hi guys, i'm looking into cloning of a big chunk of genomic sequence of about 25kb to put into a vector and transfect it into some cell line. My question is will plasmid still work or do i need to shift to bac or cosmid? how big a sequence is considered too big for plasmid? I don't really know much about BAC and cosmid and i can't seem to find out the info i need. So if anyone can also help direct me to some articles it...
  http://www.protocol-online.org/forums/topic/10293-plasmid-bac-or-cosmid/
*  Cloning Mitochondrial Targeting Sequence - Molecular Cloning - BioForum
Cloning Mitochondrial Targeting Sequence - posted in Molecular Cloning: Dear All,I'm actually having problems to clone a 95 bp fragment into a 7kbp vector.I ordered the dsDNA 95 bp oligo, containing a mitochondrial targeting sequence (MTS) from microsynth.I included the restriction sites for EcoRI and BamHI, with additional 6 bp facing outwards.Sequence:tctgcagaattcATGTCCGTCCTGACGCCGCTGCTGCTGCGGGGCTTGACAGGCTCGGCCCGGCGGCTCCCAGTGCCGCGCGCCAAGggatccaccatgUpper letters: MTSUnderlined: Rest...
  http://www.protocol-online.org/forums/topic/29143-cloning-mitochondrial-targeting-sequence/
*  Cloning of GFP - Molecular Cloning - BioForum
Cloning of GFP - posted in Molecular Cloning: Hi there, So I recently ligated my gene into a Pet32a vector using restriction sites using EcoR1 and Xho1. From another Pet vector I have to create primers for the GFP insert containing BgII and EcoR1 sites. Just wondering, once I amplify my GFP (using BgII/EcoR1 sites), how do I ligate it into my original construct..Do I cut my original constuct with EcoR1 and BgII (because looking at the vector map, Pet32a has a BgII site and ligate?) Che...
  http://www.protocol-online.org/forums/topic/23184-cloning-of-gfp/
*  Help cloning!!! - Molecular Cloning - BioForum
Help cloning!!! - posted in Molecular Cloning: Hello every body I hope someone can gimme a clue! Problem: I need to clone a fragment of PCR of about 4.2 Kb in a plasmid of about 5.3Kb. Oligos are containing 2 different ER sites: BamH1 and Xho1. BamH1 sites is 4 nucleotide away from the end (5 if you count the A), XHo is 5 or 6 far away from the end. Xho and Bam are 50 bp far away in the vector. Protocol I digest 4 ug of vector and all the PCR product with bamh1 for 6 h...
  http://www.protocol-online.org/forums/topic/7949-help-cloning/
*  Stuck from last august! need help urgently - cloning and pcr ! - Molecular Cloning - BioForum
Stuck from last august! need help urgently - cloning and pcr ! - posted in Molecular Cloning: Hi I have been trying to clone a pcr product of 3065bp in pEF6V5 His TOPO vector. I have tried everything but couldnt amplify my product with normal taq. I have tried phusion (neb), recombinant taq (fermentas, sigma, invitrogen, intron biotech), dreamtaq (fermentas), platinum taq (invitrogen) and also Ex taq from takara but in vain as none gave any result. Thought phusion (neb) gives ban...
  http://www.protocol-online.org/forums/topic/28389-stuck-from-last-august-need-help-urgently-cloning-and-pcr/
*  Cloning - Plant Molecular Biology | Sigma-Aldrich
DNA cloning or molecular cloning is the biological or laboratory process of creating genetically identical organisms, or exact copies of DNA fragments. The process of cloning a DNA plasmid involves fragmentation of the DNA vector, ligation of the new nucleotide fragment onto an appropriate cloning vector, transfection of the product into a cell, then selection of the successfully transfected cells and expression of the gene. Nucleotide sequences, or genes, can also be replicated by the polymerase chain reaction (PCR). Recombinant DNA, or chimeric DNA, is that which has been made from two different species. Ligases are used to catalyze the joining of two complementary DNA probes or single-strands of DNA. Genetic engineering involves direct manipulation of an organism′s genome, including growing cells in microbial media, examining alleles, and using expression vectors or constructs to introduce a specific gene into a target ...
  http://www.sigmaaldrich.com/life-science/molecular-biology/molecular-biology-products.html?TablePage=9434926
*  Odd point mutations - Molecular Cloning - BioForum
Page 1 of 2 - Odd point mutations - posted in Molecular Cloning: Lately I've been doing a lot of cloning into pBluescript and some derivatives made in our lab. The plasmid is then going into XL1-Blue MRF' cells by electroporation. This has been a successful and efficient strategy in the past, but lately, sequencing of the plasmid constructs has shown a seemingly inordinate amount of point mutations in the sequence. I originally asked a question regarding this a couple months ago,...
  http://www.protocol-online.org/forums/topic/9694-odd-point-mutations/
*  Subcloning into pMIR-REPORT-luc - Molecular Cloning - BioForum
Subcloning into pMIR-REPORT-luc - posted in Molecular Cloning: Hey everyone, I'm about to pull my hair out over this and am hoping to find some help here. I'm trying to use Ambion's pMIR-REPORT-luc reporter plasmid to clone in a 40bp insert in the 3' UTR of the luciferase gene. The restriction sites are SpeI and HindIII and are located 62bp from each other. I've conducted around 6 digestion, ligation and transformation experiments varying around the protocol below an...
  http://www.protocol-online.org/forums/topic/10406-subcloning-into-pmir-report-luc/
*  New Yeast Cloning System for Producing Proteins with Native Amino Acid,,,Sequences ( One-column protein purification and n...)
... One-column protein purification and native eukaryotic amino acid se... figure 1 ...Stratagenes ESP LIC cloning and expression system offers rapid ...Cloning PCR products using the LIC strategy is so efficient that as li...,New,Yeast,Cloning,System,for,Producing,Proteins,with,Native,Amino,Acid,,,Sequences,biological,advanced biology technology,biology laboratory technology,biology device technology,latest biology technology
  http://www.bio-medicine.org/biology-technology/New-Yeast-Cloning-System-for-Producing-Proteins-with-Native-Amino-Acid-0D-0A-0ASequences-44-1/
*  Digestion and ligation problem! Need advice - Molecular Cloning - BioForum
Digestion and ligation problem! Need advice - posted in Molecular Cloning: I'm trying to ligate a 700 bp fragment into a 7 kb vector- I've PCR amplified the 700 bp fragment (which was in a 3 kb vector) with restriction enzyme sites on each end (the same sites that I made my vector have in order to ligate). When I designed my primers, I added 3 extra bases to each end after the enzyme site.
  http://www.protocol-online.org/forums/topic/18760-digestion-and-ligation-problem-need-advice/
*  Molecular Cloning and Biological Characterization of a Novel Murine Lymphoid Growth Factor | JEM
The cloning of a novel murine gene that encodes a protein with lymphopoietic activity is described. The activity was initially identified in a coculture system of an adherent thymic stromal cell line with a medullary phenotype, Z210R.1, and day 15 fetal liver (6). Interestingly, the nonadherent lymphoid-like cells that grew in these cocultures phenotypically resembled B cells. A clonal lymphoid line (NAG8/7) was derived from long term cocultures, and this line expresses B220, 6C3 (BP-1), and is surface μ+, but κ chain negative. Importantly, NAG8/7 cells proliferated in response to conditioned medium from the Z210R.1 cell line and thus became the basis of a bioassay that allowed for the preliminary characterization of the growth factor and the eventual cloning of the cDNA encoding this activity.. The cDNA encoding the NAG8/7 growth activity was cloned from a library made from the Z210R.1 cell line. Based on the nucleotide sequence, the mature TSLP protein consists of 121 ...
  http://jem.rupress.org/content/192/5/671