 94 degrees C 30 seconds  64 degrees C 30 seconds  72 degrees C 1 minute  After 25 cycles finish with 72 degrees C for 5 minutes Novablue cells were.

Slides:

Advertisements

Similar presentations

Subcloning Techniques

Advertisements

JY BB JK JK CS LE MG AR NG JS SJ YW MK DS LL DH JB.

Recombinant DNA Technology

1 PCR RESULTS! Josef Lautin Erica Forsberg Zdravka Boskovic Henrik Kleinhans Adam Kundurakis Håkan Lundberg.

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

Recombinant DNA Technology. Recombinant DNA Technology combines DNA from different sources – usually different species Utility: this is done to study.

Chapter 4: recombinant DNA

Preparation of competent E. coli cells Transformation of the ligated construct into E. coli Purification of the digested fragment Purification of the pBR322.

Cloning Vector Map.

Polymerase Chain Reaction - PCR The photocopier of molecular biology.

CISC667, F05, Lec3, Liao CISC 667 Intro to Bioinformatics (Fall 2005) Molecular Biology Tools Gel electrophoresis Cloning PCR DNA Sequencing.

DNA Science Day 2 Extracting, Ligating and Transforming Physical Biology Bootcamp October 2006 Caltech.

DNA Science Day 2 Extracting, Ligating and Transforming APh162 Winter 2005 Caltech.

Cloning:Recombinant DNA

Preparation of competent E. coli cells Preparation of probes by PCR Digestion of pBR322 and plasmid with restrionenzymes BamH1, EcoR1 and EcoRV Purification.

Cloning into Plasmids Restriction Fragment Cloning & PCR Cloning by the Topo TA™ Method.

CULTURE INDEPENDENT ANALYSIS OF MICROBIAL COMMUNITIES IN SOIL

Genetic engineering to produce an organism which will make a ‘foreign’ protein:  Obtain ‘foreign’ gene  Amplify using PCR  Insert gene into a vector.

EDVOKIT#300: Blue/White Cloning of a DNA Fragment

Trends in Biotechnology

Recombinant DNA I Basics of molecular cloning Polymerase chain reaction cDNA clones and screening.

DNA Cloning and PCR.

Week 7 Wednesday: –Screening of library transformants –Innoculation of colonies for plasmid preps –Practice PCR Turn in Lab #11 Thursday: –Plasmid minipreps.

Genetic Engineering An Overview. What is it??? Applied techniques of genetics and biotechnology (“Wet lab procedure”). Much trial and error. Applied techniques.

Confirmation of positive clones Screening of positive clones Selection of high copy number clones Selection of positive clones FLUTCORE vaccine yeast constructs.

Practical training: Test for the presence of a pathogen in a wash water potato by PCR.

Unit 2-5 Notes Mr. Hefti – Pulaski Biology Genetic Testing.

The DNA structure differences between X and Y chromosomes of Silene latifolia and Y chromosomes of Silene latifolia Roman Hobza, Martina Lengerová, Pavla.

The Polymerase Chain Reaction Some milestones In molecular biology recognised by the award of the Nobel prize.

Molecular Biology II Lecture 1 OrR. Restriction Endonuclease (sticky end)

1 SURVEY OF BIOCHEMISTRY Nucleic Acids continued… Amino Acids.

Molecular Genetic Technologies Gel Electrophoresis PCR Restriction & ligation Enzymes Recombinant plasmids and transformation DNA microarrays DNA profiling.

Today Extension product purification (direct sequencing) House Keeping

Chapter 16 Microbial Genomics “If we should succeed in helping ourselves through applied genetics before vengefully or accidentally exterminating ourselves,

Molecular Cloning.

Molecular Cloning. Definitions   Cloning :   Obtaining a piece of DNA from its original source (Genome) and introducing it in a DNA vector   Sub-cloning:

Lab 22 Goals and Objectives: EDVOKIT#300: Blue/White Cloning of a DNA Fragment Calculate transformation efficiencies Compare control efficiency to cloned.

The Factor II (Prothrombin) G20210A Detection and Genotyping

SURVEY OF BIOCHEMISTRY Nucleic Acids continued… Amino Acids

Lecture 9 January 23, 2016 Biotech 3.

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

EDVOKIT#300: Blue/White Cloning of a DNA Fragment

YAC and BAC cloning systems

hisGFPmek and PDZ domain

First, convert 0.125° into minutes and seconds.

YAC and BAC cloning systems

Molecular Cloning.

Gene Isolation and Manipulation

Lab 8: PTC Polymerase Chain Reaction Lab

CISC 667 Intro to Bioinformatics (Spring 2007) Molecular Biology Tools

Small RNA Sample Preparation

mRNA Sequencing Sample Preparation

Recombinant DNA Unit 12 Lesson 2.

ChIP DNA Sample Preparation

Total uninduced vector only- Group1 2- Total uninduced spp382– Group1.

EDLC(Embedded system Development Life Cycle ).

Simulating Genetic Screening

Molecular Cloning.

1- Molecular weight marker 2- BamHI 3- BamHI 4 –EcoRI 5 – Hind III

Jung-Ok Han, Sharri B Steen, David B Roth Molecular Cell

Molecular Biotechnology

1- Molecular weight marker 2- BamHI 3- BamHI 4 –EcoRI 5 – Hind III

Use of short-term culture for identification of Mycobacterium avium subsp. paratuberculosis in tissue from Crohn's disease patients D. Schwartz, I. Shafran,

C.2.10 Sample Questions.

Volume 8, Issue 6, Pages (December 2003)

C.2.8 Sample Questions.

C.2.8 Sample Questions.

For vectors {image} , find u + v. Select the correct answer:

Genomic DNA Sample Preparation

Cell Culture Engineering XI

Presentation transcript:

 94 degrees C 30 seconds  64 degrees C 30 seconds  72 degrees C 1 minute  After 25 cycles finish with 72 degrees C for 5 minutes Novablue cells were used

Our PCR Product Molecular weight marker

Our vector after purification, it’s pretty weak but it’s there.

No colonies were formed after ligation. Therefore a backup culture made by Lasse was used for screening and analysis. Novablue cells were used

Reference PCR fragment from another group Ligated vector, pretransformation. No vector is visible. Screening of Lasses colonies, you can clearly see the result when comparing our leftmost sample with the PCR reference.