miRNA transfection using oligofectamine

Slides:

Advertisements

Similar presentations

Physiological 3D Tissue Model of the Airway Wall and Mucosa Melanie M. Choe Alice A. Tomei Melody A. Swartz Press return to continue.

Advertisements

Subcloning Techniques

Please refer to the AimPlex assay user manual for details of the assay procedure.

Cat # SL Store at 4 0 C GenJet Plus DNA In Vitro Transfection Reagent A Protocol for Transfections of Mammalian Cell 100 l 500 l 1000 l.

Extraction of Nucleic Acids (Genomic DNA, mRNA and Plasmid DNA)

Tian He Media non-selective: YPD without antibiotics Selective: Synthetic complete dropout medium (SC) (auxotroph) YPD with antibiotics.

Micron-sized particles Li Junhua Abstract Micron-sized agarose particles were prepared using emulsification/ gelation method as a reservoir.

Cloning a DNA segment from bacteriophage Recombinant DNA transformed into bacterial cells Last week we plated cells onto agar plates + ampicillin + X-gal.

Cloning a DNA segment from sheep Recombinant DNA transformed into bacterial cells Last week we plated cells onto agar plates + ampicillin + X-gal Controls:

Week 5 Wednesday: –Ligation of chromosomal digests into plasmid vectors – part II – Checking ligation success Thursday: –Electrocompetent cell preparation.

Cloning with Plasmids Genetic Engineering Invented.

Recombinant DNA Bacterial Transformation Student Instructions Transformation.

QuantityMaterialSymbol 1Late Exponential phase culture of wildtype E. coli W3110, a K12 strain 3Tryptose Blood Agar Base plates (TBAB) 2TBAB plates containing.

Cat # SL Store at 4 0 C GenJet Plus In Vitro DNA Transfection Reagent A Protocol for generation of Lentivirus from 293T cell 100 l 500 l.

Cat # SL HEPG2 Store at 4 0 C GenJet  In Vitro DNA Transfection Reagent for HepG2 Cells (Ver. II) A Protocol for Transfecting HepG2 Cells.

Cat # SL Store at 4 0 C LipoJet DNA In Vitro Transfection Reagent A Protocol for Transfections of Mammalian Cell 100 l 500 l 1000 l

Transformation with Firefly gene Lab Protocol

Cat # SL Store at 4 0 C GenJet In Vitro DNA Transfection Reagent A Protocol for Transfections of Bacmids Into Sf9 Cells 100 l 500 l 1000.

C. Elegans Unit: Lab Activities

YEAST LAB DEAD YEAST W/MINERAL OIL

Cat # SL Store at 4 0 C GenJet In Vitro DNA Transfection Reagent A Protocol for Transfections of Mammalian Cell 100 l 500 l 1000 l

Cat # SL Store at 4 0 C PolyJet In Vitro DNA Transfection Reagent A Protocol for Transfections of Mammalian Cell 100 l 500 l 1000 l

Cat # SL CHO Store at 4 0 C GenJet In Vitro DNA Transfection Reagent for CHO Cells (Ver. II) A Protocol for Transfections of CHO Cells 100.

Cellular Respiration Lab Problem: What are the products of cellular respiration and fermentation? Background: Hypothesis:

Cat # SL Store at 4 0 C SL SuperFection™ DNA Transfection Reagent Small 0.5 ml Large 1.0 ml Gaither Drive Gaithersburg, MD FAX.

Cat # SL Store at 4 0 C LipoD293 In Vitro DNA Transfection Reagent (Ver. II) A Protocol for generation of Lentivirus from 293T cell 100 l.

Cat # SL Store at 4 0 C GenJet DNA In Vitro Transfection Reagent 100 l 500 l 1000 l Gaither Drive Gaithersburg, MD FAX

MTT ASSAY Induction of Cell Proliferation by ConA.

Antibody Array Assay Report 1. Protocol 2 Protein Extraction 1.Wash the cells with ice cold 1X PBS. 2.Add Lysis Beads and Extraction Buffer to the sample.

▶ Objective : To know methods that make new cell line through stable transfection 김준섭.

Cell Culture Techniques 程洪强，方瑜，孙明姣 2013 年 12 月. Cell Thawing and Transfection.

Cat # SL Store at 4 0 C LipoD293 DNA In Vitro Transfection Reagent (Ver. II) 100  l 500  l 1000  l Gaither Drive Gaithersburg, MD

How to bioengineer a novel bio-photovoltaic system?

Add 20uL Opsonization Buffer (OB) to shaded wells 1.

Cat # SL Store at 4 0 C GenJet™ siRNA Transfection Reagent Small 0.5 ml Large 1.0 ml Gaither Drive Gaithersburg, MD FAX

M M M M 5. Not Listed

Bacterial Transformation Lab

Recombinant DNA Bacterial Transformation Student Instructions Ligation.

Lab meeting Nguyen Thi Dai Trang. Electroporation of K562, Hela, IM9  Protocol 1. 2x10 6 cells 2. PBS wash 2 times 3. Suspend in 90µl PBS.

Total bacterial number is an important indication for food quality

Cat # SL Store at 4 0 C GenJet In Vitro DNA Transfection Reagent A Protocol for Transfections of Mammalian Cell 100 l 500 l 1000 l

Protein Purification bYSY.

Introduction Recent years have seen an explosion in the number and variety of plant molecular biology applications being used in research laboratories.

Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products.

Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products.

Immunostaining 5μm Tissue Sections for Confocal Imaging

Products > CHO Transfection Reagent (Chinese Hamster Ovary Cells)

Transfection Reagent GenJet™ In vitro siRNA Small 0.5 ml Large 1.0 ml

Products > HUH-7 Transfection Reagent (Liver Cancer Cells)

Pre-Lab: pGLO Bacterial Transformation

Bacterial Transformation Visual Guide

Aim: To perform the technique of radial immunodiffusion.

Mammalian Cell l 500 l l Gaither Drive Gaithersburg, MD 20877

Practical methods BSS020X329A

Lab Activity 5 Biuret Method

Supplemental Figure 1 HA-GIPR WT Untransfected HA-C46S HA-G198C

Products > KB Transfection Reagent (Mouth Epidermal Carcinoma)

Products > HeLa Transfection Reagent (Cervix Adenocarcinoma Cells)

Lab Activity 6 Biuret Method

Volume 11, Issue 1, Pages (January 2003)

LNCaP * C4-2B Du145 22Rv1 SI Figure 1. miR-124 inhibits the proliferation of CaP cells. WST-1 cell proliferation analysis of AR-positive CaP cell lines.

pIRS1(S636/639) IRS1 actin PF Supplemental Figure S1

PTB1 nPTB Ai Aii GM03201 P1+N1 GM2132 P1+N1 GM2132 C1 GM2132 P1

Products > NCI-H292 Transfection Reagent (Lung Carcinoma, CRL1848)

The effects of solution temperature on catalase reaction rates

Products > CLBPEC Transfection Reagent (Neuroblastoma Cells)

GenJet In Vitro DNA Transfection Reagent for Hela Cells (Ver

Forensic DNA Fingerprinting:

Effects of neomycin on CVB3-Nancy replication kinetics.

Presentation transcript:

miRNA transfection using oligofectamine (Invitrogen) Day 1 : Plate 100 000 HeLa cells per well (6-well plate) Day 2 : Per well 2 µl oligofectamine + 13 µl OPTIMEM 1.7 µl miRNA (20µM solution) + 183.3 µl OPTIMEM Mix Incubate 15 min. @ room temperature In the meantime, wash cells with pre-warmed OPTIMEM once. Add 800 µl OPTIMEM per well. Add the miRNA/oligofectamine mix. Leave 4 hours @ 37°C Add 1 mL complete DMEM. Leave cells overnight Day 3 : Retransfect cells as decribed before Add 1 mL complete DMEM. Leave cells 48h Day 5 : Harvest cells.