Breeding for low-temperature stress tolerance in Hevea …

Six clones such as P 21, P 110, P 107, P 20, P 17, and P 57 were found superior to the check clones RRII 430, RRIM 600, RRII 429, and RRII 105, while clone P 17 exhibited better performance in the ...

High-throughput clone screening followed by protein …

2.1. Sample preparation for the clone screening assay. Single colony was picked up from transformed plate for each gene of interest (Table 1) and transferred in sterile tube (1.5 ml) with 50 μl distilled water followed by boiling for 5 min with a subsequent centrifugation at 11,000 g for 5 min.The clone construct in the supernatant was used as …

Harvesting and analyzing clones following CRISPR gene editing

A CRISPR-Cas9 gene engineering workflow for clonal selection – application note. The following protocol covers how to harvest and analyze your clones following …

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Development of physiological indices for screening …

Genetic material and water deficit treatment. Ninety clones of E. camaldulensis and thirty-six clones of E. tereticornis used in the present study were sourced from the breeding program executed by the Institute of Forest Genetics and Tree Breeding, Coimbatore, India. Six-month old cuttings with average height of ~ 20 cm and …

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Screening Strategies for High-Yield Chinese Hamster …

Yang et al. Screening for High-Yield Clones. cells have endogenous GS genes that can be selected by adding MSX at low levels (Bebbington et al., 1992; Brown et al., 1992). CHO-K1 cell line was first used in the GS system (Cockett et al., 1990). GS-knockout cell lines are also developed to improve the

From deep sequencing to actual clones

Here, using the isolation of antibody genes from a phage and yeast display selection as an example, we show the power of a rapid and simple inverse PCR-based method to easily …

Use of a FLAER-based WBC assay in the primary screening of PNH clones

PNH clones were detected with the FLAER assay in 63 (11.8%) of 536 samples tested, whereas PNH RBCs were detected in only 33 (6.2%), and always with a smaller clone size. The FLAER assay on WBCs is a more sensitive and robust primary screening assay for detecting PNH clones in clinical samples.

Breeding for low-temperature stress tolerance in

Identification of high yielding genotypes with abiotic stress tolerance is one of the most important aspects of crop improvement in Hevea to expand rubber cultivation to non-traditional regions and to meet the needs of the domestic market. This study was aimed to evaluate 20 clones for yield potential and cold stress tolerance and for subsequent …

A high-throughput, restriction-free cloning and screening …

Here, we address recently developed restriction-free methods in cloning, and present a more cost-efficient protocol that has been optimized to improve both …

Easy selection of recombinant clones

We have developed a polymerase chain reaction (PCR)-based procedure to facilitate the selection of recombinant clones. The insert to be cloned is ligated to an antibiotic resistance marker. The ligation product is amplified by PCR, followed by standard cloning procedure into a bacterial vector. ... eliminating the screening of the transformants ...

Screening of 19 Salix clones in effective phytofiltration potentials …

(2018). Screening of 19 Salix clones in effective phytofiltration potentials of manganese, zinc and copper in pilot-scale wetlands. International Journal of Phytoremediation: Vol. 20, No. 13, pp. 1275-1283.

Antibody Screening by Microarray Technology—Direct Identification …

The assa y type des cribed ab ove was rar ely app lied in the l iterat ure. ... at a very early stage of clone screening and thus to pick the bes t clones for a respective application.

Screening for Specific Recombinant Clones | 16

DOI link for Screening for Specific Recombinant Clones. Screening for Specific Recombinant Clones. By Lucía Irene González-Villaseñor, Mark M. Manak. Book Recombinant DNA Principles and Methodologies. Click here to navigate to parent product. Edition 1st Edition. First Published 1998. Imprint CRC Press. Pages 60.

Screening λgt Recombinant Clones by Hybridization to …

A rapid, direct method for screening single plaques of λgt recombinant phage is described. The method allows at least 106 clones to be screened per day and simplifies physical containment of recombinants.

Accelerated clone selection for recombinant CHO cells using …

Identify desirable clones at an earlier cloning stage. Assess therapeutic protein expression label-free. Eliminate use of specific antibodies for screening. Easily …

Enhanced cell density cultivation and rapid expression-screening …

Described microscale cultivation strategy can be used for rapid high-throughput screening of P. pastoris clones, media optimization, and high-throughput recombinant protein production. The knowledge gained through this work may also be applied, to other suspension cultures, with some modifications.

Selection, Screening, and Analysis of Recombinant Clones

Learn how to identify and isolate transformants and recombinant clones using various methods and techniques in gene cloning. The chapter covers selection …

Third-Generation Sequencing and Analysis of Four Complete …

A three-step PCR screening method was used to identify PLE-positive clones by screening a Rongchang pig BAC library and PacBio third-generation sequencing was performed. BLAST comparisons and other bioinformatics methods were applied for sequence analysis. Results Five PLE-positive BAC clones, designated BAC-10, BAC …

White and green screening with circular polymerase …

will not require induction for screening. We used CPEC to clone an insert into each des-tination vector (summarized in Fig. 1). We chose CPEC because it is a directional, positional, sequence independent, and ligation independent cloning protocol.1 CPEC also requires only two steps and one enzyme. In parallel, we use PCR to linea-

Top tips for screening for Cloned Inserts

For identifying positive clones from a plasmid cloning procedure, the routine of performing a mini-prep and then checking the putative clones by restriction digestion is most commonly used. Of course, if you need to screen a large number of clones, another option is a colony PCR to identify positives, followed by restriction digests to confirm….

Third-Generation Sequencing and Analysis of Four Complete …

Specific primers were then selected based on the PCR amplification results. A three-step PCR screening method was used to identify PLE-positive clones by screening a Rongchang pig BAC library and PacBio third-generation sequencing was performed. BLAST comparisons and other bioinformatics methods were applied for sequence analysis.

Clone Screening

Dedicated solutions for clone screening and single-cell isolation. Improved probability of finding rare clones of interest. Objective selection of mammalian clones and microbial …

An improved PCR strategy for fast screening of specific and …

In this study, we have developed an improved PCR screening method, which can be used for screening of clones with unwanted random integration (RI) of the rAAV genome (Figure 1B).During gene targeting by perfect homologous recombination, strand exchange occurs between the homology arms and the targeted locus (Figure 1A), …

Somatic mutant clone screening: scan for novel NASH …

A novel method of in vivo hepatocyte clonal screening for non-alcoholic steatohepatitis (NASH) target genes in mouse liver. a In a recent Cell paper by Wang and colleagues, an Adeno-associated virus (AAV) library was created and injected into mouse liver to generate mosaic liver with multiple mutant hepatocyte clones followed by …

5 screening methods every scientist should know

Listed below are 5 commonly used methods. Blue-White Screening. Blue-white screening is a widely used technique to examine successful cloning. In this method the insert is …

Selection, Screening, and Analysis of Recombinant Clones

Direct PCR screening of Pichia pastoris clones

Direct PCR screening of Pichia pastoris clones. Direct PCR screening of Pichia pastoris clones Biotechniques. 1996 Jun;20(6):980-2. doi: 10.2144/96206bm08. Authors S Linder 1, M Schliwa, E Kube-Granderath. Affiliation …

A Simplified Method for Gene Knockout and Direct Screening …

These phenotypes therefore allow the direct screening of recombinant clones. Further, the identification of the recombinant clones can be performed directly on the transformation plates by a simple assay. In contrast to other screening possibilities commonly employed for positive differentiation of recombinants, starch clearance plates …

Comprehensive clone screening and evaluation of fed-batch …

Clone screening was conducted similarly to the proposal in Invitrogen's guide and as applied often in literature [31,36-40]: Clones were grown in the selected media, ... Ind Eng Chem Proc Des Dev. 1979; 18:357–364. doi: 10.1021/i260071a001. [Google Scholar]

Semi-continuous scale-down models for clone and …

Semi-continuous scale-down models for clone and operating parameter screening in perfusion bioreactors ... These scale-down models are very useful to design and scale-up optimal bioreactor operating conditions as well as screening for different media and cell lines. Keywords: cell culture; perfusion; scale -down ...

Phylogenetic Screening of Ribosomal RNA Gene …

clones without requiring direct DNA sequencing. Using this approach, 35 phylogenetic groups (previously iden tified by cultivation or PCR-based rRNA gene surveys) were detected and quantified among the BAC clones. Since the probability of recovering chimeric rRNA gene sequences in large insert BAC clones was low, we used

Genotyped functional screening of soluble Fab clones …

Clone selections was done based signal to background (S/B) ratio of screening immunoassay by dividing each clones time-resolved fluorescence signal with mean signal from empty controls of the same ...

Screening for resistant clones of

Screening for resistant clones of Casuarina equisetifolia to bacterial wilt and the analysis of AFLP markers in resistant clones S. Sun. ... The results showed that clones FJ11 and FJ13 were highly resistant to BW with the lowest disease incidences of 10.8 and 15.8%, respectively, whereas clones FJ9 and GD15 were the most …

Screening for Clones

Once E. coli is transformed with a ligation mixture, the task of identifying the desired clone follows. Depending on the DNA source, tens to hundreds of thousands of clones may be screened before the target is identified. As we explored in Chapter 7, the strategy used in the cloning process will determine the amount of work required to identify that clone.