FR3 Molecular Resources

Molecular Resources List

FR3 Next Generation Sequencing Service

Is an opportunity to collaborate with the FR3 scientists at Smith College on your research project, and to gain hands on experience with the Illumina MiSeq. For more detailed information about our sequencing service, click here.

Filarial RNA and DNA preparations

Filarial RNA and DNA preparations are now available through BEI Resources. If you are not currently registered with BEI, fill out the BEI Resources Material Transfer Agreement and Registration forms, then locate the FR3 catalog to order FR3 materials through BEI.

Species	Preparation	Life Cycle Stage(s)
Brugia malayi	total RNA	adult female adult male L3 L4 microfilariae
B. pahangi	total RNA	adult female adult male microfilariae
Dirofilaria immitis	total RNA	adult female adult male L3 microfilariae
Aedes aegypti	total RNA	eggs male pupae young adult adult male adult female
B. malayi	genomic DNA	—–
B. pahangi	genomic DNA	—-
D. immitis	DNA	—–
Acanthocheilonema viteae	DNA	—–
Aedes aegypti	DNA	—–

PCR primers available through BEI Resources

Filarial PCR primers are now available through BEI Resources. If you are not currently registered with BEI, fill out the BEI Resources Material Transfer Agreement and Registration forms, then use the FR3 ordering link on the BEI homepage to get started.

LIBRARIES MUST BE ORDERED FROM BEI LABORATORIES (www.beiresources.org). OTHER cDNA LIBRARIES ARE ORDERED THROUGH FR3 MOLECULAR DIVISION.

Entire libraries can still be ordered through FR3 molecular division. Fill out the BEI Resources Material Transfer Agreement and Registration, then use the ‘FR3 Ordering‘ link on the BEI homepage to get started.

Characteristics of the Onchocerca volvulus Microfilaria cDNA Library. Please click each library to enter its data sheet.

*Onchocerca* *volvulus*	Microfilarial cDNA Library (SAW98MLW-OvMf)* L2 cDNA Library (SAW98MLW-OvL2)* Infective L3 cDNA Library (SAW94WL-OvL3)* Adult female cDNA Library (SAW98MLW-OvAF)* Adult male cDNA Library (SAW98MLW-OvAM)* Adult female ivermectin treated cDNA Library (SAW99PF-OvAF) Genomic library (OvG) Molting L3 cDNA Library (Ov3M)*
Onchocerca ochengi	Infective L3 cDNA Library (SAW98MLW-OoL3)
Brugia malayi	Microfilarial cDNA library (SAW94LS-BmMf) L2 cDNA library (JHU96SL-BmL2) Infective L3 cDNA library (SAW94WL-BmL3) L3 molting-day 6 larval cDNA library (SAW96MLW-BmL3d6) L3 molting-day 9 larval cDNA library (SAW96MLW-BmL3d9) L3 subtracted cDNA library (SAW97LS-BmL3SA) L3 subtracted cDNA library (SAW97YG-BmL3SB)* L3 subtracted cDNA library (SAW97LS-BmL3SD)* L4 cDNA library (SAW99MLW–BmL4)* Young adult cDNA library (SAW99MLW-BmYA)* Adult male cDNA library (SAW94NL-BmAM)* Adult female cDNA library (SAW96MLW-BmAF)* Adult cDNA library (JHU96SL-BmA) Genomic library (BmG)
Wuchereria bancrofti	Microfilarial cDNA Library (SAW95SJL-WbMf)* Infective L3 cDNA Library (SAW96MLW-WbL3) Adult female cDNA Library (JHU94SL-fWb) Adult male cDNA Library (JHU94SL-mWb)

Library Name	SAW98MLW–OvMf
Key:	SAW	Lab: SAW = Steven A. Williams’ Lab JHU = Johns Hopkins University – Alan Scott’s Lab
	98	year constructed
	MLW	initials of the individual who constructed the library
	Ov	species: Ov = Onchocerca volvulus Oo = Onchocerca ochengi Bm = Brugia malayi Wb = Wuchereria bancrofti
	Mf	stage: Mf = microfilaria L2 = second larval stage L3 = third larval stage L3d6 = L3 to L4 molt, day 6 L3d9 = L3 to L4 molt, day 9 L3SA (L3SD) = L3 cDNA subtracted with microfilaria, adult male and adult female cDNA L3SB = L3 cDNA obtained by subtracting JHU93SL-BmL3 cDNA library (constructed using SL sequence and PCR) from SAW94WL-BmL3 cDNA library (constructed using conventional cDNA methods). L4 = fourth larval stage YA = young adult AF = adult female AM = adult male

GENOMIC LIBRARIES

malayiBAClibrary clones – used to map the Wolbachia genome (see Foster et al. 2004. Int. J. Parasit. 34: 733-746; and Foster et al. 2001. BioTechniques 30(6): 1216-1222). Please contact Smith College (genome@smith.edu) for ordering information.

FILARIAL SERUM SAMPLES/ANTIGENS

TO ORDER FILARIAL SERUM SAMPLES AND ANTIGENS, YOU MUST HAVE A CURRENT FR3 REGISTRATION FORM ON FILE, AND A SMITH COLLEGE MTA.

Bm14 Antigen for serologic monitoring

Positive Control for the Binax Filariasis Now test

Filarial Serum Bank

BmWSP MONOCLONAL ANTIBODY – THIS MONOCLONAL IS AVAILABLE THROUGH BEI LABORATORIES (www.beiresources.org).

Fill out the BEI Resources Material Transfer Agreement and Registration, then use the FR3 Ordering link on the BEI homepage to get started. This monoclonal was developed by Patrick Lammie at Centers for Disease Control, and recognizes wBm0432.

Real-time PCR Assay for the Simultaneous Detection of Wuchereria bancrofti and Brugia malayi

When conducted in accordance with the published protocol [1], this multiplex real-time PCR assay is capable of detecting Wuchereria bancrofti and Brugia malayi DNA simultaneously, or independently, at sensitivities which parallel the published singleplex assays for parasite detection. This assay has been tested on worm extracted DNA, as well as DNA extracted from both human bloodspots and vector mosquito pools. Xenomonitoring studies conducted in co-endemic locations can benefit from the reagent-cost and labor-cost related savings that this assay is able to provide.

1). Pilotte N, Torres M, Tomaino FR, Laney SJ, Williams SA. A TaqMan-based multiplex real-time PCR assay for the simultaneous detection of Wuchereria bancrofti and Brugia malayi. Mol Biochem Parasitol 2013 May; 189(1-2): 33-37.

Smith College will provide primers, controls and protocol for this procedure.

Please contact (genome@smith.edu) for further information regarding this assay.

FR3 Brugia malayi list of putative T cell epitopes

A list of putative T cell epitopes identified using the iVax Immunoinformatics Toolkit, by screening over 11,000 proteins belonging to the genome-derived B. malayi protein sequences (EpiVax, Providence, Rhode Island), in collaboration with Dr. Anne De Groot at the University of Rhonde Island.

Brugia malayi putative T cell epitopes