in vitro transcription and translation protocols

in vitro transcription and translation protocols

... in vitro RNA synthesis. In vitro translation of a targeted message has been shown to be inhibited in the presence of antisense RNA in vitro (14), and in vivo translation has been blocked in ... polynucleotide kinase. 15. Incubate kinase reaction for 30 min at 37OC. 16. Heat inactivate the polynucleotide kinase by incubating the reaction at 70°C for 30 min. 17....

Ngày tải lên : 11/04/2014, 09:47

422
268
0

Báo cáo khoa học: The inﬂuence of cold shock proteins on transcription and translation studied in cell-free model systems pdf

... translational level, the system of combined transcription ⁄ translation was decoupled. The in uence on the transcriptional level was investigated by in vitro transcription of the CAT gene using ... BcCSP. In vitro translation For determining the in uence of CSPs on the transcription apparatus the system for combined transcription ⁄ translation was used without T7 RNA p...

Ngày tải lên : 07/03/2014, 21:20

12
487
0

Báo cáo khoa học: In vitro selection and characterization of a stable subdomain of phosphoribosylanthranilate isomerase potx

... for 10 min. In vitro selection for mAb M2 binding was by adding 200-lL aliquots of the supernatant to the selection matrix and incubating at room temperature for 30 min. Unselected protein–plasmid ... the re-introduction of an initiator methionine codon and was achieved by PCR with primers incorporating the necessary sequence. The product was re-inserted into pMS401, and the resulti...

Ngày tải lên : 30/03/2014, 20:20

14
382
0

Báo cáo y học: " Preparation of RGD-modified Long Circulating Liposome Loading Matrine, and its in vitro Anti-cancer Effects"

... USA) containing 10% heat-inactivated fetal bovine serum (Nuoding, China), and the HT-29 cells were grown in DMEM medium (GIBCO, USA) also containing 10% heat-inactivated fetal bovine serum. ... saline (PBS), and resuspended at 2 × 10 6 cells /mL in Annexin-V binding buffer. The supernatant (100 μL/tube) was incubated with 5 μL of Annexin-V-FITC (Invitrogen, USA) and 5 μL of...

Ngày tải lên : 26/10/2012, 08:57

12
635
0

Analysis, sequencing and in vitro expression of PCR products

... functional domain mapping of proteins where fragments that represent protein domains are expressed and functionally analyzed using in vitro assays. Furthermore, rapid in vitro protein–protein interaction studies ... with Analysis, sequencing and in vitro expression of PCR products 89 Analysis, sequencing and in vitro expression of PCR products 5.1 Introduction Analysis...

Ngày tải lên : 25/10/2013, 22:20

24
494
0

Tài liệu Báo cáo khoa học: Proteolysis of Pseudomonas exotoxin A within hepatic endosomes by cathepsins B and D produces fragments displaying in vitro ADP-ribosylating and apoptotic effects doc

... in the mitochondrial apoptotic response induced by the toxin remains to be determined. Finally, we assign an important role to the endosomal acidic cathepsins B and D in increasing the in vitro ... cathepsins B and D, with the major cleavages generated in vitro occurring mainly within domain III of ETA-A. Cell-free endosomes preloaded in vivo with ETA intraluminally proces...

Ngày tải lên : 18/02/2014, 04:20

15
588
0

Tài liệu Báo cáo khoa học: The endogenous retinoid metabolite S-4-oxo-9-cis-13,14-dihydro-retinoic acid activates retinoic acid receptor signalling both in vitro and in vivo pdf

... gentamicin (Gibco Invitrogen), 1% (v ⁄ v) l-glutamine, 5 lgÆmL )1 insulin (Gibco Invitrogen), 10 ngÆmL )1 epidermal growth factor (Gibco Invitrogen) and 240 lgÆmL )1 Geneticin Ò (G418; Gibco Invitrogen). ... (Gibco Invitrogen), 1% (v ⁄ v) l-glutamine (Gibco Invitrogen) and 1% (v ⁄ v) pyruvate (Gibco Invitrogen). Mouse mammary epi- thelial cells (HC11) were grown in RPMI 1640+ medium (Gi...

Ngày tải lên : 18/02/2014, 08:20

17
406
0

Tài liệu Báo cáo khoa học: Regulation of connective tissue growth factor (CTGF/CCN2) gene transcription and mRNA stability in smooth muscle cells Involvement of RhoA GTPase and p38 MAP kinase and sensitivity to actin dynamics docx

... ed-end actin and determine the level and distr ibution o f F -actin [34]. Therefore, it i s tempting to speculate that interactions between monomeric G-actin and actin-binding proteins are a ... fixed, permeabilized and stained for F-actin with rhodamine-conju- gated phalloidin. Th e effects of toxin B and latrunculin B on the actin filaments was examined by preincubating the cells wit...

Ngày tải lên : 19/02/2014, 16:20

15
576
0

Tài liệu Báo cáo khoa học: Template requirements and binding of hepatitis C virus NS5B polymerase during in vitro RNA synthesis from the 3¢-end of virus minus-strand RNA docx

... radioactivity incorporated into the nucleic acids was measured after TCA precipitation and plotted against the incubation time in minutes. T. Astier-Gin et al. Binding and replication of 3¢-end of HCV minus ... of the HCV minus-strand RNA is illustrated in Fig. 1. This fragment contains two domains: domains I and II. Structures of both domain I (A), as determined by Schuster et al...

Ngày tải lên : 20/02/2014, 01:20

15
597
0