Abstract：
:Studies on the regulation of human milk protein genes and suitable cell culture systems have been limited due to restricted availability of tissue samples from lactating women. Although mammary gland tissue has been available from mammary reduction surgery, studies on tissue-specific expression of milk protein genes require samples obtained during specific stages of expression in the tissue, i.e., lactation. We have therefore developed a technique to isolate mRNA and genomic DNA directly from epithelial cells isolated from human milk and used PCR methodology to specifically amplify the cDNA and genomic DNA for human beta-casein. When comparing fresh human milk with milk that has been stored for five hours at 37 degrees C or frozen and thawed, we found that the amount of mRNA isolated was considerably higher from fresh milk. However, in spite of this, we could still isolate intact mRNA from frozen milk. The described methodology should be useful in studies on milk protein gene expression during lactation and studies on variants of milk protein genes.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Lindquist S,Hansson L,Hernell O,Lönnerdal B,Normark J,Strömquist M,Bergström Ssubject
Has Abstractpub_date
1994-10-01 00:00:00pages
692-6issue
4eissn
0736-6205issn
1940-9818journal_volume
17pub_type
相关文献
BIOTECHNIQUES文献大全abstract：:The recently developed ultrasensitive reverse transcriptase (RT) test involving a PCR step can detect minute amounts of RT in single retroviral particles and is 10(6) times more sensitive than conventional RT assays. We have found that different DNA-dependent DNA polymerases like DNA Polymerase I from Escherichia coli...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1996-02-01 00:00:00
abstract：:Monitoring morphogenetic processes, at high resolution over time, has been a long-standing goal of many developmental cell biologists. It is critical to image cells in their natural environment whenever possible; however, imaging many warm-blooded vertebrates, especially mammals, is problematic. At early stages of dev...
journal_title：BioTechniques
pub_type：
doi：10.2144/03342st01
更新日期：2003-02-01 00:00:00
abstract：:We have developed a high-throughput, multiplex reverse transcription PCR (RTPCR) assay that is suitable for the analysis of medium-to low-copy cellular RNA transcripts from small numbers of cells (10(4)). High throughput was attained by utilizing microplate-based RNA extraction and RTPCR protocols, followed by PCR pro...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/97226st02
更新日期：1997-06-01 00:00:00
abstract：:S-palmitoylation (S-acylation) is emerging as an important dynamic post-translational modification of cysteine residues within proteins. Current assays for protein S-palmitoylation involve either in vivo labeling or chemical cleavage of S-palmitoyl groups to reveal a free cysteine sulfhydryl that can be subsequently l...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/000114516
更新日期：2017-02-01 00:00:00
abstract：:Reporter assays that use luciferase are widely employed for monitoring cellular events associated with gene expression. In general, firefly luciferase and Renilla luciferase are used for monitoring single gene expression. However, the expression of more than one gene cannot be monitored simultaneously by this system b...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/05386ST03
更新日期：2005-06-01 00:00:00
abstract：:We report a simple and inexpensive method to quantitate loading and transfer of RNA and to examine RNA integrity for use with Northern hybridization. This technique involves quantitation by computer-assisted video densitometry of a negative photograph of 28S and 18S rRNA from ethidium bromide-stained RNA fixed to nylo...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1992-02-01 00:00:00
abstract：:Complete brain fixation can be achieved with transthoracic cardiac infusion without thoracotomy. Light and electron microscopy tissue sections reveal preservation of cytoplasmic and nuclear structure at all magnification levels. Punched samples were obtained from the fixed tissue specimens in precisely localized areas...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/000112003
更新日期：2005-10-01 00:00:00
abstract：:Plasmid pUC18rspL is a 3.788-kilobase pair vector, derived from pUC18, pKK232-8 and pHSG664, which identifies promoter-bearing DNA fragments functional in StrA E. coli by activation of a promoterless streptomycin-sensitive gene cartridge (rspL). Expression of the plasmid-borne rspL gene leads to sensitivity dominance ...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1990-12-01 00:00:00
abstract：:In this special section of BioTechniques, we examine the role of rapid molecular technologies in the detection and identification of agents of infectious disease (ID) and biological weapons (BWs). Besides the threat posed by the global proliferation of BW technologies, there are numerous emerging and reemerging ID age...
journal_title：BioTechniques
pub_type： 杂志文章,评审
doi：10.2144/03354ss01
更新日期：2003-10-01 00:00:00
abstract：:The concentration of proteins in cells is an important parameter that determines how a protein will interact with other proteins or pharmacological agents. Recent developments in Western blotting techniques have now made this a method of choice to measure protein concentration in complex solutions such as total cell e...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1999-03-01 00:00:00
abstract：:Polyethyleneimine (PEI) is a flocculent that is widely used in the downstream purification of monoclonal antibodies. It is an in-process residual that is carried through the drug purification process and strongly inhibits residual DNA quantitation by real-time quantitative PCR assay. Very high sample dilutions (e.g., ...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/btn-2020-0011
更新日期：2020-06-01 00:00:00
abstract：:The Sequence Analysis Workshop (SAW) is an interactive program for sequence analysis of immunoglobulin variable domains. Sequences for SAW can be obtained from GenBank or from a standard text file. SAW can compare a variable domain to as many as 100 different sequences, calculate the extent of homology, sort the seque...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1993-12-01 00:00:00
abstract：:A modification of the TRI Reagent procedure has been elaborated for isolation of RNA from polysaccharide- and proteoglycan-rich material. In the modified procedure, RNA is precipitated from the aqueous phase by the combined action of isopropanol and a high-salt concentration. Under these conditions, RNA is effectively...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1995-12-01 00:00:00
abstract：:[Formula: see text] Known to be a sturdy weapon in a scientist's arsenal, how has the gene editing tool CRISPR been applied in the fight against COVID-19? ...
journal_title：BioTechniques
pub_type： 新闻
doi：10.2144/btn-2020-0145
更新日期：2020-11-01 00:00:00
abstract：:Tissue engineering scaffolds have emerged as a powerful tool within regenerative medicine. These materials are being designed to create environments that promote regeneration through a combination of: (i) scaffold architecture, (ii) the use of scaffolds as vehicles for transplanting progenitor cells, and/or (iii) loca...
journal_title：BioTechniques
pub_type： 杂志文章,评审
doi：10.2144/000113754
更新日期：2011-10-01 00:00:00
abstract：:Cell-imaging approaches using new laser-based technologies have a wide applicability to thefields of pathology and cell biology. Here, we present the application of several of these techniques, including confocal scanning laser microscopy (CSLM), laser scanning cytometry (LSC), and laser capture microdissection (LCM),...
journal_title：BioTechniques
pub_type： 杂志文章,评审
doi：10.2144/01314rv01
更新日期：2001-10-01 00:00:00
abstract：:Sarah Webb explores how RNA sequencing innovations help scientists tackle the complexity of the transcriptome. ...
journal_title：BioTechniques
pub_type： 新闻
doi：10.2144/btn-2017-0105
更新日期：2018-02-01 00:00:00
abstract：:Gel retardation analysis, or band shift assay, is technically the simplest method to investigate protein-nucleic acid interactions. In this report, we describe a nonradioactive band shift assay using a fluorescent DNA target and an ALFexpress automatic DNA sequencer in place of the current method that utilizes radioac...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/01305rr03
更新日期：2001-05-01 00:00:00
abstract：:We have developed a novel vector pTCS, as a tool for efficient selection of open reading frame (ORF)-containing inserts. In pTCS clones containing an insert with an ORF a downstream marker gene (immE3, conferring resistance to colicin) is activated via translational coupling with the insert, and transformed cells can ...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/000112629
更新日期：2007-12-01 00:00:00
abstract：:Metabolic and electrical coupling through gap junction channels is implicated in cell differentiation, tissue homeostasis, and electrotonic propagation of signals in excitable tissues. The characterization of gating properties of these channels requires electrophysiological recordings at both single- and multiple-chan...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/03345rr03
更新日期：2003-05-01 00:00:00
abstract：:We describe a new real-time fluorescence video microscope design for capturing intensified images of cells containing dual wavelength "ratio" dyes or multiple dyes. The microscope will perform real-time capture of two separate fluorescence emission images simultaneously, improving the time resolution of spatial distri...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1990-03-01 00:00:00
abstract：:A pipeline has been created for the characterization of Arabidopsis thaliana mutants by generating flanking sequence tags (FSTs) and optimized for economic, high-throughput production. The GABI-Kat collection of T-DNA mutagenized A. thaliana plants was used as a source of independent transgenic lines. The pipeline inc...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/03356st01
更新日期：2003-12-01 00:00:00
abstract：:There is growing consensus on the potential use of pharmacogenetics in clinical practice, and hopes have been expressed for application to the improvement of global health. However, two major challenges may lead to widening the "biotechnological gap" between the developing and the industrial world;first the unaffordab...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：2005-10-01 00:00:00
abstract：:We describe a method using an inexpensive craft glue to routinely isolate specific areas of tissue as small as 1 mm2 from paraffin sections. The tissue may be digested to release nucleic acid suitable for PCR or reverse transcription PCR. The use of this procedure obviates the requirement for manual microdissection or...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/96205st04
更新日期：1996-05-01 00:00:00
abstract：:Piezoelectric dispensing of proteins from borosilicate glass capillaries is a popular method of protein biochip fabrication that offers the advantages of sample recovery and noncontact with the printing substrate. However, little regard has been given to the quantitative aspects of dispensing minute volumes (1 nL or l...
journal_title：BioTechniques
pub_type：
doi：10.2144/03342mt02
更新日期：2003-02-01 00:00:00
abstract：:An improved method was developed for in situ reverse transcription-polymerase chain reaction (RT-PCR) to detect and localize mRNA in tissue sections. The coverslip mounted-immersion cycled (COSMIC) in situ RT-PCR technique combines the advantages of solution-phase PCR with the tissue immobilization necessary for in si...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/98241st02
更新日期：1998-01-01 00:00:00
abstract：:A nonimmune phagemid recombinant antibody fragment (rFab) library was generated with a nominal diversity of 1.16 x 10(7) using the QuikChange Multi Site-Directed Mutagenesis kit. Two degenerate primers spanning the third complementarity-determining region (CDR) loops of the antibody fragment light and heavy chain were...
journal_title：BioTechniques
pub_type：
doi：10.2144/03354st05
更新日期：2003-10-01 00:00:00
abstract：:SOP3 is a web-based software tool for designing oligonucleotide primers for use in the analysis of single nucleotide polymorphisms (SNPs). Accessible via the Internet, the application is optimized for developing the PCR and sequencing primers that are necessary for Pyrosequencing. The application accepts as input gene...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/05381RR01
更新日期：2005-01-01 00:00:00
abstract：:We describe a simple and inexpensive method of performing sequencing reactions for 24 single-strand M13 DNA clones in microtiter plates. To simplify elevated temperature incubations during sequencing reactions, two heating blocks were designed to accommodate microtiter plates and fit within common laboratory heating m...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1990-07-01 00:00:00
abstract：:The Drosophila heart has gained considerable traction as a model of cardiac development and physiology. Previously we described a semiautomatic optical heartbeat analysis (SOHA) method for quantifying functional parameters from the fly heart that facilitated its use as an organ system and disease model. Here we presen...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/000114255
更新日期：2015-02-01 00:00:00
