BAC SEQUENCING REQUEST FORM
===========================

Fields marked with * are required fields.

* BAC NAME:  
* LIBRARY: 
   ESTIMATED SIZE OF THE BAC: 
   BAC END SEQUENCE: 	SP6 end:
			T7 end:
			Extra sequences:
* WHAT IS THE ANCHORING MARKER:
* WHERE DOES THE ANCHORING MARKER MAP TO:
	* Marker type:
	* Chromosome:
	   Offset (genetic distance from the top of the chromosome): 
	   Confidence (LOD SCORE):
	   If PCR based marker, what are the primers:
	* Population mapped:
	* Polymorphism:
	   If f2.2000, what is the mapping data:
	   If there is a picture file for mapping, please attach it:
	   Is there a sequence for this marker:

IS THERE ANOTHER MARKER ASSOCIATED WITH THIS BAC:
	* Marker type:
	* Chromosome:
	   Offset:
	   Confidence (LOD SCORE):
	   If PCR based marker, what are the primers:
	* Population mapped:
	* Polymorphism of L. esculentum (PCR size etc):
	   If f2.2000, what is the mapping data:
	   If there is a picture file for mapping, please attach it:
	   Is there a sequence for this marker:

 * WAS THE MAPPING POSITION OF THE BAC RECONFIRMED: 
	If yes, which method did you use:
		PCR amplification
		Southern hybridization
		Mapping RFLP marker developed from  BAC end sequence
	Is there any picture for this: 
	
   IS THERE A DIGESTION PICTURE FOR THE BAC:
   IS THERE A CONTIG FOR THE BAC:
	Contig fingerprinting picture:

   WAS THE BAC TESTED FOR BEING A SINGLE CLONE:
   OTHER COMMENTS:
	

How to confirm the BAC as a single clone without contamination:
	Streak the BAC culture on the LB plates with 12.5 ug/ml chloramphenichol.  After growing 
overnight, randomly pick 10 clones and do the miniprep.  One method is to run PFG (pulsed field gel) 
gel for NotI digested DNA together with the lamda ladder PFG marker (New England Biolab, #340).  If 
all ten clones have the same insert size, then there is only one BAC in the culture or glycerol 
stock.  Then prepare a stab in the 1.5 ml tube and send to us.  The alternative method for this is 
to run HindIII digested DNA on a regular agarose gel slowly overnight.  If ten clones have the same 
digestion pattern, then there is no contamination problem for this clone.  
THE FIRST METHOD IS PREFERRED, SINCE THE SIZE OF INSERT CAN BE ESTIMATED.  

How to prepare a stab in the 1.5 ml tube:
	Pour about 1ml of LB+agar medium with 12.5 ug/ml chloramphenichol into 1.5 ml tube.  After 
solidification, stab the toothpick (dipped into the fresh culture) in the LB agar.  Grow the tubes 
overnight and ship to us at room temperature. 

Mailing address: 

Attn: Eileen Wang
240 Emerson Hall
Dept of Plant Breeding
Cornell University
Ithaca, NY 14853