User Panel

My Panel

Bookmark Science Articles

insert Protocols

Search results for: insert

Protocols » Search Results

Search Results Protocol Links Sort by: Hits | Alphabetical

Additional slide pre-treatment for mapping smaller insert clones - http://www.sanger.ac.uk/HGP/methods/cytogenetics/additional_treatment.shtml

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Additional slide pre-treatment for mapping smaller insert clones for FISH. Sanger Institute. - [Read Additional slide pre-treatment for mapping smaller insert clones]

Amplification of Insert End Sequences from BACs Clones by Thermal Asymmetric Interlaced PCR - http://pubs.nrc-cnrc.gc.ca/ispmb/ispmb16/16175-2.pdf

Category: BACs Bacterial Artificial Chromosome Protocols

Protocol presents the amplification of insert end sequences from bacterial artificial chromosome clones using TAIL-PCR. The amplified products are suitable as probes for chromosome walking and genome mapping and as templates for direct sequencing. The protocol has been used in rice genome studies. - [Read Amplification of Insert End Sequences from BACs Clones by Thermal Asymmetric Interlaced PCR]

Bacteriophage Lamda Plant Genomic Library Construction Protocol - http://www.oardc.ohio-state.edu/stockingerlab/Protocols/GenomicDNAPhageLibraryConstruction.pdf

Category: DNA Protocols: Genomic DNA Library Protocols

Protocol for bacteriophage lamda plant genomic library construction. Includes: Preparation of the insert DNA; Pilot Sau3A Digestion; Full-scale partial digestions. - [Read Bacteriophage Lamda Plant Genomic Library Construction Protocol]

C. elegans RNAi Protocol - http://www.zoology.ubc.ca/~alorch/rnai-protocol.htm

Category: C. elegans Protocols: C. elegans Genetics Protocols

Protocol for C. elegans RNAi. Includes: Transformation of competent cells; Blunt-end ligation; Preparation of competent cells; Dephosphorylation of linear plasmid DNA; Restriction Digest: EcoRV; Insert amplification from gDNA; Gel purification: QiaQuick gel purification kit; Mini-prep; Transformant Screening; Bacterial preparation and induction; Preparation of worms for RNAi feeding. - [Read C. elegans RNAi Protocol]

Cloning into Bacteriophage M13 Vectors Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4018

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

Protocol describes three standard methods to construct bacteriophage M13 recombinants: (1) ligating insert DNA to a linearized vector, prepared by cleavage of M13 RF with a single restriction enzyme; (2) using alkaline phosphatase to suppress self-ligation of the linearized vector, and (3) using M13 RF cleaved with two restriction enzymes for directional cloning. - [Read Cloning into Bacteriophage M13 Vectors Protocol]

Colony PCR Protocol II - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4141

Category: PCR Protocols: Colony PCR

Protocol describes the use of PCR to screen for bacteria that carry recombinant plasmids. The PCR can be carried out using the same primers as for amplification of the cloned insert. To determine the orientation of the insert, a third, insert-specific primer that is asymmetrically distanced from the clonal insertion site can be used. - [Read Colony PCR Protocol II]

Construction of the Sequencing Library Protocol - http://sequence-www.stanford.edu/protocols/library.html

Category: DNA Protocols: DNA Sequencing Protocols

Protocol first describes the vector preparation and, then, describes the insert preparation. Vital to have an excellent vector in order to produce a sequencing library. Protocol employs the male-specific coliphage M13 as the sequencing vector. M13 is a filamentous phage with a single-stranded, circular genome. M13 is widely used as a vector because many versions are available commercially and because M13 has certain advantages. - [Read Construction of the Sequencing Library Protocol]

Cycle Sequence Reactions For Large Insert Plasmid Templates Protocol - http://bacpac.chori.org/cyclesere.htm

Category: DNA Protocols: DNA Sequencing Protocols

Protocol for cycle sequence reactions for large insert plasmid templates. Includes: CycleSequenceConditions; Primers Useful For BAC and PAC Vectors. - [Read Cycle Sequence Reactions For Large Insert Plasmid Templates Protocol]

Direct Sequencing Using MultiPlex PCR-Based Method - http://www.genome.ou.edu/MultiplexPCRbasedSeq.html

Category: DNA Protocols: DNA Sequencing Protocols

Direct sequencing from amplified bacterial and large insert cloned human or mouse genomic DNA via an improved MultiPlex PCR-based method. Includes: Preparing primers for MP-PCR; Amplification; PCR Product Clean-up; Sequencing. - [Read Direct Sequencing Using MultiPlex PCR-Based Method]

DNA Cloning - http://www.molecularstation.com/dna-cloning/

Category: Molecular Biology Protocols: DNA Ligation Protocols

DNA Cloning. Cloning Calculator for ratio insert to vector / plasmid. Molecular Station. - [Read DNA Cloning]

DNA Insert Ligation into Vector DNA (with T4 DNA Ligase) - http://www.fermentas.com/techinfo/modifyingenzymes/protocols/p_dnainslig.htm

Category: Molecular Biology Protocols: DNA Ligation Protocols

Using digested vector DNA (50-400ng) and foreign DNA to be inserted. Fermentas - [Read DNA Insert Ligation into Vector DNA (with T4 DNA Ligase)]

DNA Ligation - http://www.bio.com/protocolstools/protocol.jhtml?id=p2053

Category: Molecular Biology Protocols: DNA Ligation Protocols

Description and protocol of the steps required to join together plasmid and insert fragments to create a new plasmid. Paul Kaufman, Univ California. - [Read DNA Ligation]

DNA ligation with Linearized DNA - http://www.genome.ou.edu/protocol_book/protocol_partII.html#II.D

Category: Molecular Biology Protocols: DNA Ligation Protocols

DNA ligations are performed by incubating DNA insert with linearized cloning vector in the presence of ligation buffer, rATP, and T4 DNA ligase. Roe. Univ. Oklahoma. - [Read DNA ligation with Linearized DNA]

EMBL Cloning protocols - http://www.embl-hamburg.de/display?file=~geerlof/webPP/protocoldb/Cloning/pdb_cloning.html

Category: DNA Protocols: DNA Cloning

Preparation of oligo solutions PCR experiments, Digestion of insert DNA, Digestion and dephosphorylation of vector DNA, Ligation of DNA fragments with sticky ends, Ligation of DNA fragments with blunt ends, Preparation of chemically competent E. coli - [Read EMBL Cloning protocols]

Identification of End Clones in YAC Subclone Libraries Protocol - http://humgen.wustl.edu/hdk_lab_manual/yeast/yeast10.html

Category: Yeast Protocols: Yeast Nucleic Acid Protocols: Yeast PCR Protocols

To identify the YAC subclones containing both a human insert and a portion of either the left or right arm of the pYAC4 vector. Identification of these clones is necessary in order to do YAC chromosome walking, and is also useful in the determination of whether a particular YAC clone has a contiguous human insert or whether a co-cloning event has occurred. Vector arm sequences are identified using pBR322 fragments from a BamHI-PvuII double digest. - [Read Identification of End Clones in YAC Subclone Libraries Protocol]

Ligation Optimization Protocol - http://www.fhcrc.org/science/labs/fero/Protocols/Ligation.html

Category: Cloning Protocols

Protocol can be used to optimize ligation conditions for difficult to clone (e.g. very large) fragments. The principle is to independently characterize the ligation kinetics of the vector and insert DNA fragments and then to combine them in optimal ratios. - [Read Ligation Optimization Protocol]

Natural History Museum Protocol for EST sequencing from lambda zap phage plaques - http://www.nhm.ac.uk/hosted_sites/schisto/protocols/nhm_est_method.html

Category: RNA Protocols: cDNA Libraries Library

(by PCR of phage insert, cycle sequencing using ABI dye terminator kits and automated sequencing on an ABI 373A) Ian Ridgers - [Read Natural History Museum Protocol for EST sequencing from lambda zap phage plaques]

Subcloning of Yeast Artificial Chromosomes into Phage Lambda Protocol - http://humgen.wustl.edu/hdk_lab_manual/yeast/yeast9.html

Category: Yeast Protocols: Yeast Genetics Protocols: YACs Protocols

Protocol for the subcloning of Yeast artificial chromosome into phage lambda. To subclone the large insert fragment of human DNA contained within a YAC into a bacteriophage lambda vector. The subclones are 15 to 23 kb in size, and can be used to identify new polymorphic markers from a known region of the genome, to map a specific locus, and/or to screen other libraries. - [Read Subcloning of Yeast Artificial Chromosomes into Phage Lambda Protocol]

Articles (1-6 of 6)

Lambda Phage Protocol Large DNA Preparation

Single Stranded Plasmid DNA Isolation Protocol

A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.

Antibodies Phage Expression Protocol Using 96-well Microtiter Plates

A Phage display expression protocol for antibody expression in 96-well microtiter plates.

Constructing Random-Peptide Libraries for Phage Display

The protocol describes the construction of a large (10^8 to 10^10) primary random peptide phage library in fUSE5 or f88-4 strains.

DNA Ligation Protocol

The DNA Ligation protocol described here contains the steps required to join together using ligase enzyme both plasmid DNA and insert DNA fragments in order to create a new plasmid. This new ligated plasmid can be transformed after into competent bacteria to produce DNA for mini, midi or maxi-prep isolation.

Vector Design Protocol for Transgenic Mice Generation

The protocol gives general considerations for the design of targeting vectors for transgenic mice. The protocol shares tips in the design of knock-out and knock-in vectors and some of their strategies for producing homologously recombined embryonic stem cells.

[1-6]