L 50.2 52.0 54.1 56.5 59.4 62.3 65.2 68.5 72.4 1000 bp 500 bp 250 bp Annealing Temperature in Degrees C V4 16s – 506f-Y/806rB ITS2 – ITS3NGS_mix/ITS4NGS.

Slides:

Advertisements

Similar presentations

PCR Techniques. Basics of PCR Primers –15-60bp (60bp is limit synthesized by IDT) –Annealing temp ideally >55C (portion that anneals to your template)

Advertisements

Recipe for ??????????????????????????????? What you will make Ingredients What to do

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.

PCR way of copying specific DNA fragments from small sample DNA material "molecular photocopying" It’s fast, inexpensive and simple Polymerase Chain Reaction.

Detection of the human Mitochondrial DNA A Polymerase Chain Reaction Experiment.

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

How PCR works Cold Spring Harbor Animation Animated .GIF files

Working summary Wang Qian. Analyse the 16SrDNA fragment by DGGE The samples come from China. DNA extraction : use the Wizard Genomic DNA Purification.

FISH 543 / OCEAN 575 Molecular Techniques

Virginia iGEM Workshop #4 High School Education Series.

1. Sample & Primer CEDDGP001- 카톨릭의대 정의달 * 총 10 개 샘플중에서 4, 14, 24, 28, 36 샘플에 대해서만 PCR 실험을 수행하였다. PCR 실험 수행시 DNA 농도를 100ng/ul 로 희석하여 사용하였다.

PCR of our OV What is PCR again? I forgot… Preparing for PCR Running a PCR.

Davidson Day Ateh Neuroscience Centre, Institute of Cell and Molecular Sciences Barts and The London School of Medicine and Dentistry Queen Mary University.

PCR and qRT-PCR May 3. PCR The thermocycle analyzing the products essential components of the reaction optimization basic rules of primer design problems.

Polymerase Chain Reaction (PCR)

PCR POLYMERASE CHAIN REACTION Dauphin Island Graduate Neurobiology.

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

Polymerase Chain Reaction. Invented by Kary Mullis Mullis and Faloona, Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction.

PCR Troubleshooting Virginia Balke

Chapter 14: DNA Amplification by Polymerase Chain Reaction.

Insect DNA Barcoding. Agenda DNA Bug Barcoding at FLCC Finger Lakes Invertebrate Biodiversity Study (FLIBS) Introduction to Biology (for majors), Research.

PCR Forensics. Today’s Lab There has been an outbreak of Salmonella poisoning in the Student Union cafeteria at Stanford University cafeteria. You have.

Polymerase Chain Reaction (PCR) Developed in 1983 by Kary Mullis Major breakthrough in Molecular Biology Allows for the amplification of specific DNA fragments.

POLYMERASE CHAIN REACTION (PCR) Bridges Polymerase Chain Reaction  Simple reaction  Produces many copies of a specific fragment of DNA  Live.

Success criteria - PCR By the end of this lesson we will be able to: 1. The polymerase chain reaction (PCR) is a technique for the amplification ( making.

V IRAL Q UEST Lesson 8: Reverse Transcription. Reverse Transcription  Reverse Transcription converts RNA into complementary DNA.  This DNA strand can.

Polymerase Chain Reaction (PCR)

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

The polymerase chain reaction

The polymerase chain reaction

Amplification of a DNA fragment by Polymerase Chain Reaction (PCR) Ms. Nadia Amara.

Detection of the human VNTR using PCR* *A Polymerase Chain Reaction Experiment.

DNA sequencing. How DNA synthesis takes place inside the living cell?

PCRPCR Presented by : Rana AL-Turki. 1- Definition of PCR. 2- Requirements for PCR. 3-PCR Process. 4-Procedure.

Introduction to PCR Polymerase Chain Reaction

PUC 19 5’ 3’ 5’ 3’ 5’ 3’ 5’ 3’ PCR -I pUC 19 specific primers Amplicon purification PCR -II 5’ 3’ 5’ 3’ 5’ 3’ 5’ 3’ 5’ 3’ 5’ 3’ Composite pUC 19 primers.

HRM REAL TIME PCR Presented by: Dadkhah Fahimeh SNP genotyping by HRM REAL TIME PCR.

The Factor II (Prothrombin) G20210A Detection and Genotyping

PCR mediated mutagenesis 2013 년도 2 학기 생화학 실험 (2) 5 주차 조교 : 안성원.

Polymerase Chain Reaction (PCR): DNA Replication in vitro DNA replication in vivo DNA replication in vivo –Occurs in S-phase –Helicase, primers, DNA polymerase.

Lecturer: Bahiya Osrah Background PCR (Polymerase Chain Reaction) is a molecular biological technique that is used to amplify specific.

HRM Assay and Optimization 1. DNA Quality 2. Amplicon LengthAmplicon  lengths of 100–300 bp 3. Primer Selection 4. Dye Selection 5. MgCl2 Concentration.

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.

I. PCR- Polymerase Chain Reaction A. A method to amplify a specific piece of DNA. DNA polymerase adds complementary strand DNA heated to separate strands.

Presented by: Khadija Balubaid.  PCR (Polymerase Chain Reaction) is a molecular biological technique  used to amplify specific fragment of DNA in vitro.

Introduction to PCR Polymerase Chain Reaction

NJSDGP003_부미경 PCR 1. Sample list 2. Primer list.

Success criteria - PCR By the end of this lesson we will be know:

Polymerase Chain Reaction (PCR)

PCR conditions: 94ºC for 15 min

IGEM Bioware Summer Week 7!.

Today’s Title: CW: DNA manipulation – separating and probing

(B.P :51) ( B:P52 ).

Experimenttal part.

qPCR experiment design

Oligose - Primers Qiagen Dneasy Polymerase Chain Reaction What Does it all Mean? Maria Brown October 22, 1009.

Presented by : Mostafa al-mosavi

Gradient PCR using Taq polymerase with 125ng primer concentration

Polymerase Chain Reaction & DNA Profiling

PCR How does PCR work?: Separation of two strands

Polymerase Chain Reaction (PCR)

أهداف المحاضرة مراجعة المفاهيم والعلاقات الأساسية لتحليل العلاقة بين التكاليف والحجم والربح. بناء نموذج لتحليل التعادل في حالة وجود منتج وحيد. بناء نموذج.

Introduction to Polymerase Chain Reaction (PCR)

Restriction Enzyme Digestions

Detection of Genomic Variations in BRCA1 and BRCA2 Genes by Long-Range PCR and Next-Generation Sequencing Imma Hernan, Emma Borràs, Miguel de Sousa Dias,

Martin J. Somerville, Kathleen A. Sprysak, Mark Hicks, Basil G

The polymerase chain reaction

' · · ,.-.., '' !'",. -,..._ ·-.·-...;.· -

POLYMERASE CHAIN REACTION

Polymerase Chain Reaction (PCR)

Presentation transcript:

L 50.2 52.0 54.1 56.5 59.4 62.3 65.2 68.5 72.4 1000 bp 500 bp 250 bp Annealing Temperature in Degrees C V4 16s – 506f-Y/806rB ITS2 – ITS3NGS_mix/ITS4NGS Test Sample = 29 Clear choice? No Difference? Gradient PCR using degenerate bacterial/archaeal primers (V4 region 16 – 506f-Y/806rB) and fungal primers (ITS2 region – ITS3NGS_mix/ITS4NGS). Primers contain Nextera Adapters. PCR Recipe: Thermocycler Program Cyc1 (1X) 98oC for 2 min Cyc2 (35X) 98 oC for 10 s Gradient (50-72 oC) for 10 s 72 oC for 15 s Cyc3 (1X) 72 oC for 5 min Cyc4 4 oC HOLD For one 10 ul reaction mixture: Ingredient Stock Conc. ul per sample Final Conc. Invtirogen Platinum SuperFi Master Mix 2 X 5 1 515f-Y (or ITS3NGS_mix) 10 uM 0.5 0.500 806rB (or ITS4NGS) Template (FxxB2) ng/uL Sterile PCR H2O 3 Total rxn volume =