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Sample Submission Guidelines — Morrisville

For Gene Expression, Genotyping (US), Biologics, all GLP/CLIA Projects and Other Services.

Express Sequencing

Sample Type	Minimum Sample Requirements
Plasmid DNA or Purified PCR Products	500 ng per sequencing reaction at 50 ng/µl in 10 µl of water
Plasmid DNA and Primer Mixed	500 ng per sequencing reaction at 50 ng/µl - DNA 8 pmol Primer at 0.8 pmol/µl - Primer in a total volume of 10 µl
Primer	8 pmol per sequencing reaction at 0.8 pmol/µl in 10 µl

Regulatory Submission Sequencing

Sample Type	Minimum Sample Requirements
Bacterial Samples	Low Copy- 1-2 mls of 109-1010cells/ml High Copy- 1-2 mls of 108-109 cells/ml
Yeast Samples	1-2 mls of 108 cells/ml
Mammalian Samples	1-2 ml of 107 cells/ml

Research Grade Sequencing

Sample Type	Minimum Sample Requirements
Plasmid DNA	300-500 ng per sequencing reaction (minimum concentration 100ng/µl)   DNA as a dried down pellet, or in molecular biology grade water or elution buffer, NOT in TE.
PCR Product	20 ng for each 100 bp of amplicon per sequencing reaction (PCR Products must be single band and purified, additional charges will incur for enzymatic purification or gel extraction)

High Throughput Sequencing (96-well format)

Sample Type	Minimum Sample Requirements
Bacterial Samples	Glycerol stock-HT (~108 cells/ml) 100 µl/ sample
DNA	10µl at 100ng/µl per well

Sequencing — All Types

Sample Type	Minimum Sample Requirements
Stab or streak cultures	1 stab or plate
Viral DNA	generally 300 ng of DNA per sequencing reaction (exact amount will be included in quotation)
BAC clone	generally 300 ng of DNA per sequencing reaction (exact amount will be included in quotation)
RNA	2 µg minimum in water, TE or elution buffer
Primer	10 µl of 8 pmol/µl dilution of the primer per sequencing reaction (50 ng/ul for a 20-mer)  Please note that not all PCR primers are optimum for sequencing.  Primers should be approximately 20 bases long and the melting temperature should be between 50-65 degrees Celsius.

Shotgun Library Construction

Sample Type	Minimum Sample Requirements
Purified viral particles	1-2 mls of 1012 pfu/ml
DNA	20 µg
Stab or plates	1 stab or plate
Glycerol Stock	100 µl (~108  cells/ml)

Genetic Stability Testing

Sample Type	Minimum Sample Requirements
Southern Blot-Copy Number Southern Blot-Insertion Number Southern Blot-Structural Map	This amount will vary depeding on the digest number and the blot set up.  Please see your quotation for materials required. Mammalian  = 20 µg of DNA/digest Yeast and Bacteria 5-10 µg DNA/digest
Phage Testing Plasmid Loss	Bacterial 1ml (108 cells/ml)
Northern Blot Restriction Enzyme Analysis	Bacterial 1ml (108 cells/ml) Yeast 1 ml (107 cells/ml) Mammalian 1ml (106 cells/ml) (wet cell pellet - at least 3 grams or 100 ml)

Quantitative PCR

Sample Type	Minimum Sample Requirements
Cell Banks - Bacterial	1-2 ml (108 cells/ml)
Cell Banks - Yeast	1-2 ml (107 cells/ml)
Cell Banks - Mammalian	1-2 ml (106 cells/ml)
Tissue	10-25 mg
Whole Blood	1 ml
DNA	500 ng
RNA	500 ng

GMO Testing

Sample Type	Minimum Sample Requirements
PCR Method	500 ng of young leaf per sample
Southern Method	3 g of young leaf per sample

Genotyping

Sample Type	Minimum Sample Requirements
PCR/Agarose Gel method	50-100 ng per PCR reaction
SNP & STR	custom, please call Beckman Coulter Genomics for further information

Other Services

Sample Type	Minimum Sample Requirements
Subcloning	custom, please call Beckman Coulter Genomics for further information
Northern Blot Restriction Enzyme Analysis	Bacterial 1ml (108 cells/ml) Yeast 1 ml (107 cells/ml) Mammalian 1ml (106 cells/ml) (wet cell pellet - at least 3 grams or 100 ml)