scholarly journals High Throughput Methods for Gene Identification, Cloning and Functional Genomics Using the GeneTAC™ G3 Robotics Workstation

2002 ◽  
Vol 7 (5) ◽  
pp. 65-71
Author(s):  
Rifat Hamoudi ◽  
Sally Johnston ◽  
George Hutchinson ◽  
John D'Errico
Keyword(s):  
Functional Genomics ◽  
High Throughput ◽  
Gene Function ◽  
Genomic Sequence ◽  
Gene Identification ◽  
Robotic Platform ◽  
Ann Arbor ◽  
Set Up ◽  
Entire Genomic Sequence

Using a single robotic platform, the GeneTAC™ G3, we have automated most of the processes involved in the cloning and characterisation of novel disease causing genes by addressing the following; firstly, identifying the BACs of interest and making shotgun libraries. Secondly, automating the set up of sequencing reactions using methodology that eliminates the need for DNA preparation of 384 clones. Thirdly, generating sub-libraries using selective re-arraying of library clones to enable the determination of the entire genomic sequence of the gene. Fourthly, determining gene function by combination of differential screening and mini Northerns using microarrays printed using the GeneTAC™ G3 system and hybridised using the GeneTAC™ HybStation (Genomics Solutions, Ann Arbor, USA).

scholarly journals Rapid Hypothesis Testing with Candida albicans through Gene Disruption with Short Homology Regions

1999 ◽  
Vol 181 (6) ◽  
pp. 1868-1874 ◽  
Author(s):  
R. Bryce Wilson ◽  
Dana Davis ◽  
Aaron P. Mitchell
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Candida Albicans ◽  
Gene Function ◽  
Gene Disruption ◽  
Genomic Sequence ◽  
Selectable Marker ◽  
Genome Project ◽  
Gene Products ◽  
Pcr Products

ABSTRACT Disruption of newly identified genes in the pathogen Candida albicans is a vital step in determination of gene function. Several gene disruption methods described previously employ long regions of homology flanking a selectable marker. Here, we describe disruption of C. albicans genes with PCR products that have 50 to 60 bp of homology to a genomic sequence on each end of a selectable marker. We used the method to disrupt two known genes,ARG5 and ADE2, and two sequences newly identified through the Candida genome project,HRM101 and ENX3. HRM101 and ENX3are homologous to genes in the conserved RIM101 (previously called RIM1) and PacC pathways ofSaccharomyces cerevisiae and Aspergillus nidulans. We show that three independenthrm101/hrm101 mutants and two independentenx3/enx3 mutants are defective in filamentation on Spider medium. These observations argue that HRM101 andENX3 sequences are indeed portions of genes and that the respective gene products have related functions.

Respirometry for determination of the influents SS-concentration

1996 ◽  
Vol 33 (1) ◽  
pp. 311-323 ◽  
Author(s):  
A. Witteborg ◽  
A. van der Last ◽  
R. Hamming ◽  
I. Hemmers
Keyword(s):  
Experimental Design ◽  
Activated Sludge ◽  
Substrate Concentration ◽  
Full Scale ◽  
Activated Sludge Process ◽  
Respiration Rates ◽  
On Line ◽  
Large Measurement ◽  
Set Up

A method is presented for determining influent readily biodegradable substrate concentration (SS). The method is based on three different respiration rates, which can be measured with a continuous respiration meter which is operated in a cyclic way. Within the respiration meter nitrification is inhibited through the addition of ATU. Simulations were used to develop the respirometry set-up and decide upon the experimental design. The method was tested as part of a large measurement programme executed at a full-scale plant. The proposed respirometry set-up has been shown to be suitable for a semi-on-line determination of an influent SS which is fully based on the IAWQ #1 vision of the activated sludge process. The YH and the KS play a major role in the principle, and should be measured directly from the process.

Determination of trace concentrations of copper by FIA-FAAS after preconcentration on chelating sorbents

1989 ◽  
Vol 54 (7) ◽  
pp. 1785-1794 ◽  
Author(s):  
Vlastimil Kubáň ◽  
Josef Komárek ◽  
Zbyněk Zdráhal
Keyword(s):  
Sampling Frequency ◽  
Copper Determination ◽  
Flow Rates ◽  
Sample Injection ◽  
Injection Flow ◽  
Chelating Sorbents ◽  
Set Up ◽  
Trace Concentrations ◽  
Better Than

A FIA-FAAS apparatus containing a six-channel sorption equipment with five 3 x 26 mm microcolumns packed with Spheron Oxin 1 000, Ostsorb Oxin and Ostsorb DTTA was set up. Combined with sorption from 0.002M acetate buffer at pH 4.2 and desorption with 2M-HCl, copper can be determined at concentrations up to 100, 150 and 200 μg l-1, respectively. For sample and eluent flow rates of 5.0 and 4.0 ml min-1, respectively, and a sample injection time of 5 min, the limit of copper determination is LQ = 0.3 μg l-1, repeatability sr is better than 2% and recovery is R = 100 ± 2%. The enrichment factor is on the order of 102 and is a linear function of time (volume) of sample injection up to 5 min and of the sample injection flow rate up to 11 ml min-1 for Spheron Oxin 1 000 and Ostsorb DTTA. For times of sorption of 60 and 300 s, the sampling frequency is 70 and 35 samples/h, respectively. The parameters of the FIA-FAAS determination (acetylene-air flame) are comparable to or better than those achieved by ETA AAS. The method was applied to the determination of traces of copper in high-purity water.

Extra-Column Effects in Determination of Rate Parameters by the Chromatographic Method

1996 ◽  
Vol 61 (6) ◽  
pp. 844-855 ◽  
Author(s):  
Olga Šolcová ◽  
Petr Schneider
Keyword(s):  
Chromatographic Method ◽  
Dynamic Method ◽  
Axial Dispersion ◽  
Transport Parameters ◽  
Porous Particles ◽  
Loop Detector ◽  
Single Pellet ◽  
Set Up ◽  
Extra Column Effects

It was shown that the sampling loop, detector and connecting elements in the chromatographic set-up for determination of transport parameters by the dynamic method significantly influence the response peaks from columns packed with porous or nonporous particles. A method, based on the use of convolution theorem, was developed which can take these effects into account. The applicability of this method was demonstrated on the case of axial dispersion in a single-pellet-string column (SPSR) packed with nonporous particles. It is possible to handle also responses from columns packed with porous particles by a similar procedure.

A single NGS‐based assay covering the entire genomic sequence of the DMD gene facilitates diagnostic and newborn screening confirmatory testing

2021 ◽  
Vol 42 (5) ◽  
pp. 626-638
Author(s):  
Babi R. R. Nallamilli ◽  
Alka Chaubey ◽  
C. A. Valencia ◽  
Leah Stansberry ◽  
Andrea M. Behlmann ◽  
...  
Keyword(s):  
Newborn Screening ◽  
Genomic Sequence ◽  
Confirmatory Testing ◽  
Dmd Gene ◽  
Entire Genomic Sequence
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