cDNA Synthesis Service

Slides:

Advertisements

Similar presentations

Saliva sample collection with using Oragene®DNA kit DNA extraction & purification Gel electrophoresis Experimental Outline DNA quality and quantity assessment.

Advertisements

1 st Strand Synthesis in Reverse Transcription AAAAAAAA 3’ N6 TTT TTTTT 5’ 5’ 3’ Random primer Oligo(dT) primer Sequence specific primer 1 st strand cDNA.

09/07/ 24 (yy/mm/dd) Experimenter Kashima, Takashima, Hibino, Miyazawa Note-taker Takashima Title Flow Chart Sample Date Sample NameEnzymepurification.

1 Vol. 01. p Vol. 01. p Vol. 01. p.20.

1 Vol. 03. p Vol. 03. p Vol. 03. p.21.

1 Vol. 02. p Vol. 02. p Vol. 02. p.19.

1 Vol. 03. p Vol. 03. p Vol. 03. p.16.

1 Vol. 02. p Vol. 02. p Vol. 02. p.30.

1 Vol. 03. p Vol. 03. p Vol. 03. p.35.

1 Vol. 02. p Vol. 02. p Vol. 02. p.10.

1 Vol. 01. p Vol. 01. p Vol. 01. p.14.

Molecular cloning overview Steps to prepare vector hour overnight culture 2.Minipreps (day immediately following O/N) (use Qiaprep spin miniprep.

Analysis of gene expression by real-time PCR RNA Isolation from tomato.

What if we want to know what allele(s) of beta-globin an individual has?

 Used plasmid DNA, [pDNA] = 0.2ug/uL  Made two 1% Agarose Gel: 1 with EtBr & 1 without EtBr  Prepared 6 samples of pDNA to be loaded in each gel with.

Learning Goals identify common equilibrium constants, including Keq, Ksp, Kw, Ka, Kb and write the expressions for each solve problems related to equilibrium.

C DNA SYNTHESIS Yaprak Dönmez December, % agarose, 70 V, 90 min RNA Ladder W3 G2 F2 4 EN/DA F/ITTH3 EC2 B/A 1 Y RNA Ladder.

Flow of Genetic Information:. Translation  Initiation Recruitment of mRNA to the ribosome  Elongation Synthesis of nacent polypeptide  Termination.

Practical #2: Extraction of genomic DNA from E.Coli Practical #3: Agarose Gel Electrophoresis Bertrand Ong Chan JianPeng Salanne Lee.

RNA Electrophoresis. Broad and Long Term Objective To characterize the expression of ribulose 1-5 bisphosphate carboxylase oxygenase and chlorophyll AB.

Gene Expression RNA Isolation from Emiliania huxleyi.

Basic Procedures for DNA analysis I) DNA isolation & purification: –Sample: nucleated cells –Principle: A- PURIFICATION STEPS: 1.Cell lysis 2.Removal of.

Quantifying Sample DNA. Definition Quantifying DNA: a technique to calculate the quantity (weight) of DNA (deoxyribonucleic acid) in a sample. Using a.

Final Presentation Sample Preparation Nextera TruSeq RiboZero Strand Specific Clontech smRNA 16s.

Progress Report 10/7/14. Projects Expression of iGluRs in Xenopus Oocyte Color Choice Behavior Assay – Generation of Positive Control Lines.

Laboratory: Unit 3: purify PCR product (55-65) Lecture: DNA sequencing In-Class Writing: discuss abstracts (pages 68, 157) & AEM 63: , 1997 Hand.

SPECTROPHOTOMETRY. Determines concentration of a substance in solution by Measures light absorbed by solution at a specific wavelength by using spectrophotometer.

DNA concentration and purity determination

NYKRCP004 RT- PCR report PCR Condition Temp.TimeCyclesFinal Reaction vol. 45 ℃ 30min1 50ul 95 ℃ 15min 1 95 ℃ 30sec ℃ 30sec 72 ℃ 40sec.

A Novel Approach to Identifying Differential Gene Expression Alexander Richardson Robert Pignolo.

A Novel Approach to Identifying Differential Gene Expression Robert J. Pignolo and Oriana Galardi-Este.

Laboratory: Unit 4: PCR for T-RFLP (pages 83-84) Lecture: Terminal Restriction Fragment Length Polymorphism (T-RFLP) Analysis In-Class Writing: peer review.

09/07/ 17 (yy/mm/dd) Experimenter Kashima,Shihoya Note-taker Kashima Title Flow Chart Sample Date Sample NameEnzymepurification KitProductStorage 1.Miniprep.

Department of Biotechnology Life Science Experiments 생 명 과 학 실 험생 명 과 학 실 험 특성화학부 생명공학과 식물분자생물학 실험실 6. PCR, Enzyme digestion SUBJECT 6. PCR, Enzyme digestion.

RT-PCR analysis 생화학 실험 2 조교 : 이 선 민 내선 7699, 첨단과학기술관 201-B 호 신과학원 S438 호.

Library QA & QC Day 1, Video 3

M Gradient PCR using Taq polymerase with 125ng primer concentration M: 1 kb ladder 1: 60°C 2: 62°C 3: 64.4°C 4:66.8°C 5: 69.2°C 6:71.5°C 284D.

분자생물학실험 SUBJECT RNA Isolation cDNA synthesis.

Estimation of quantity and quality of isolated DNA

Restriction Analysis of Plasmid DNA

Restriction Enzyme Digestion of Phage DNA

ExTRacting DNA from C. elegans

Invest. Ophthalmol. Vis. Sci ;45(6): doi: /iovs Figure Legend:

By: Ashneet Biln and Carling Gales

Supplemental Figure 2. (A) AtplaIVA-1 and AtplaIVA-2 null transcription lines for AtPLAIVA mRNA. RNAs from the relevant wild type Col were isolated.

Fosmid Library Construction Report

Supplementary figure 3 a b M 1 2 Probe: Unc5b 7 kb

Experimenttal part.

Gradient PCR using Taq polymerase with 125ng primer concentration

Teagasc/APC Sequencing Facility

DNA Technology.

Small RNA Sample Preparation

mRNA Sequencing Sample Preparation

Protein Synthesis A brief overview Purpose? Key Players?

Transcription -The main purpose of transcription is to create RNA from DNA because RNA leaves the nucleus to carry out its functions but DNA does not -A.

Novel Molecular Method for Detection of Bovine-Specific Central Nervous System Tissues as Bovine Spongiform Encephalopathy Risk Material in Meat and Meat.

Digital Gene Expression – Tag Profiling Sample Preparation

Molecular Biology Working with DNA.

Restriction Enzyme Digestions

C.2.10 Sample Questions.

C.2.8 Sample Questions.

Altered Gene Expression in Melanocytes Exposed to 4-Tertiary Butyl Phenol (4-TBP): Upregulation of the A2b Adenosine Receptor1 Fan Yang, Thomas L. Brown,

Thing / Person:____________________ Dates:_________________

C.2.8 Sample Questions.

BETONLINEBETONLINE A·+A·+

Gel electrophoresis of measles virus cDNA amplicons (HU2 control measles virus RNA) amplified under optimised reaction conditions. Gel electrophoresis.

Agarose gel electrophoresis of rRNA after repression of r-protein synthesis. Agarose gel electrophoresis of rRNA after repression of r-protein synthesis.

Supplementary Fig. 1 bp E F G H I Hybrid 1 SP2/0 MW P3X (-)

15 - Stairways and Ladders

Presentation transcript:

cDNA Synthesis Service Date: 2007. 10.01 cDNA Synthesis Service Source of RNA 1-1. Sample Information: M1, 갯지렁이 Total RNA 1-2. Total RNA Concentration: NanoDrop® ND-1000 UV-Vis Spectrophotometer Tube # Name Conc. (ng/ul) A260/A230 A260/A280 Total vol. (ul) Total quantity 1 M1 Total RNA -1 639.05 0.96 1.85 1200 ~ 0.766mg 2 M1 Total RNA-2 690.62 0.85 1.83 ~ 0.828mg 3 갯지렁이 Total RNA-1 2244.52 1.44 2.01 1000 ~ 2.244mg 4 갯지렁이 Total RNA-2 2425.09 1.42 2.00 ~ 2.425mg Electrophoresis of RNA 1 2 3 4 5 Lane Sample name Quantity 1 Positive Control 1.0 g 2 M1 Total RNA -1 3 M1 Total RNA-2 4 갯지렁이 Total RNA-1 5 갯지렁이 Total RNA-2 28S RNA (4.8 kb) 28S RNA (4.8 kb) 18S rRNA (1.8 kb) 18S rRNA (1.8 kb) 3. mRNA Isolation Tube # Name Conc. (ng/ul) A260/A230 A260/A280 Total vol. (ul) Total quantity 1 M1 Total RNA 53.02 1.69 1.85 30 ~1.590 ug 2 갯지렁이 Total RNA 948.47 2.55 2.06 ~28.454 ug - 1 / 2 -

cDNA Synthesis Service Date: 2007. 10.01 cDNA Synthesis Service 4. Synthesis of ds cDNA Tube # Name Conc. (ng/ul) A260/A230 A260/A280 Total vol. (ul) Total quantity 1 M1 Total RNA 70.30 2.37 1.70 50 ~ 3.515 ug 2 갯지렁이 Total RNA 402.51 2.56 1.88 ~ 20.125 ug 5. Electrophoresis of ds cDNA 1kb Ladder 1 2 3 4 Lane Sample name Quantity 1 Positive Control 50ng 2 1kb Ladder 3 M1 Total RNA 100ng 4 갯지렁이 Total RNA 200ng - 2 / 2 -