摘要:
Methods are provided herein for identifying rare and/or unknown DNA sequences by next-generation sequencing approaches. Isolated double-stranded (ds), single-stranded (ss), or ds/ss DNA is fragmented and the fragments are polished, phosphorylated, and tailed, as necessary. Fragmentation can be enzymatic or mechanical. A universal adapter sequence is ligated to each fragment, wherein the adapter can have a top strand without a 5′ phosphate, a 3′ with an —H in place of the —OH, and/or a 3′ extra base complementary to any base added to the polished fragments. The ligatamers may then serve as templates for amplification using a forward primer complementary to the adapter sequence and a reverse primer targeted to the fragment sequence. Compositions produced by these methods and kits adapted for performing these methods are also described herein.
摘要:
Compositions and methods for remodeling complex populations of microbes are provided herein. RNA-guided nuclease systems are engineered to target sites in chromosomal DNA of a targeted prokaryotic, wherein the level of targeted prokaryote can be modulated in a mixed population of prokaryotes.
摘要:
The present invention provides RNA-guided endonucleases, which are engineered for expression in eukaryotic cells or embryos, and methods of using the RNA-guided endonuclease for targeted genome modification in in eukaryotic cells or embryos. Also provided are fusion proteins, wherein each fusion protein comprises a CRISPR/Cas-like protein or fragment thereof and an effector domain. The effector domain can be a cleavage domain, an epigenetic modification domain, a transcriptional activation domain, or a transcriptional repressor domain. Also provided are methods for using the fusion proteins to modify a chromosomal sequence or regulate expression of a chromosomal sequence.
摘要:
A composition, method and device for the preparation of biological samples for subsequent instrumental analyses, such as GC, GC-MS, LC and LC-MS analysis, using a solid phase extraction (SPE) process is described. Through SPE process alone or an integrated combination of protein precipitation, filtration, and SPE using a hydrophobic zirconia-coated chromatographic media, interfering compounds, such as proteins, glycerides and phosphate-containing compounds, are eliminated from the biological, food, environmental and biotechnology samples, affording an enhanced analyte response during the instrumental analysis.
摘要:
The present invention provides cell lines deficient in mannosyl (alpha-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase I (Mgat1). Also provided are methods for producing the Mga1 deficient cell lines and methods for using the Mgat1 deficient cell lines for the production of recombinant proteins having simple glycoforms.
摘要:
The present disclosure provides genetically engineered cell lines comprising chromosomally integrated synthetic sequences having predetermined epigenetic modifications, wherein a predetermined epigenetic modification is correlated with a known diagnosis, prognosis or level of sensitivity to a disease treatment. Also provided are kits comprising said epigenetically modified synthetic nucleic acids or cells comprising said epigenetically modified synthetic nucleic acids that can be used as reference standards for predicting responsiveness to therapeutic treatments, diagnosing diseases, or predicting disease prognosis.
摘要:
The present invention provides cell lines deficient in mannosyl (alpha-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase I (Mgat1). Also provided are methods for producing the Mga1 deficient cell lines and methods for using the Mgat1 deficient cell lines for the production of recombinant proteins having simple glycoforms.
摘要:
Molybdenum complexes and use thereof in thin film deposition, such as CVD and ALD are provided herein. The molybdenum complexes correspond in structure to Formula (I) and Formula (II), wherein R1, R3, R5, R7, R8 and R10 are independently and at each occurrence alkyl; R2, R6 and R9 are independently alkyl; R4 and R11 are independently and at each occurrence selected from the group consisting of alkyl, alkenyl, and alkynyl; x, z, a, c, d and f are independently zero, 1, or 2; y, b and e are independently zero or 1; and n and m are independently zero to 5.
摘要:
The present disclosure provides compositions and methods for acetylating histones at targeted chromosomal locations in a cell. In particular, the disclosure provides a fusion protein comprising a DNA binding domain and at least one histone acetyltransferase (HAT) domain, such that the DNA binding domain targets the fusion protein to a targeted chromosomal location and the HAT domain acetylates histones at the targeted location.