BDGP: Laboratory Methods

In Situ Hybridization Using Digoxigenin Labeled Probes
The BDGP Cytogenetics Core uses this protocol to localize P1s, BACs, and P element insertion sites on the polytene chromosome map.

cDNAs & ESTs

96-well RNA In Situ Hybridization Protocol
This is the protocol used to generate the embryonic whole-mount in situ expression patterns. It is an adaptation of standard protocols to a 96-well format, and includes PCR protocols for pBS and quantification of RNA.

Protocols Used to Isolate Membrane-Bound Polysomes and to Construct the Normalized CK cDNA Library
Isolation of mRNA from Drosophila rough endoplasmic reticulum
Normalization of a Drosophila cDNA library by hybridization to gDNA beads

PCR protocol for genomic DNA

Sizing cDNA PCR products
This protocol gives the PCR method used to size cDNA clone inserts.

pOT2 Vector

pOTB7 Vector

pOTB7D Vector

pFLC-I Vector

Expression Vectors

In-Fusion Protocol
Method for cloning full-length ORFs using the Clontech PCR cloning system

PCR Amplification of cDNAs fromBacterial Cultures: DGC/pOT2

Large Inserts: PCR Amplification of DGC cDNAs greater than 3.5kb

PCR Amplification of D. melogaster UniGene Collection
Using plasmid DNA as template

Transposon Insertions

Inverse PCR & Cycle Sequencing of P Element Insertions for STS Generation
These are the protocols we use in our insertional mutagenesis projects for determining the genomic DNA sequence adjacent to the point of insertion of a P transposable element, thereby allowing us to map the precise site of the element's insertion. The most updated protocols allow a single skilled technician to process 96 fly lines per week from frozen flies to DNA sequence.

This page last updated on: 9/22/05

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