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Use nanodrop to measure concentration of pEntry vectors, make 5 femtomolar working solution of each pEntry.

Excel sheet set up to calculate the necessary volumes

LR (concentration) calculations.xlsx

Sample calculation:

DNADNA length from Geneious (kb)Conc (ng/ul)Final Vol of working solution (ul)Conc of working solution (fM) Vol of DNA to add, V (ul)Vol EB (ul)
 lxyf 0.66*y*l*f/x 
pENTR_LilrB24.42001005 7.2692.74
pENTR_PirB5.12001005 8.41591.585
pENTR_Hef1a3.92001005 6.43593.565
pDEST7.920010010 26.0773.93

Combine into 1 aliquot:

  • 1uL of 5 fmol of pENTR_L4_Promoter_R1
  • 1uL of 5 fmol pENTR_L1_Gene_L2
  • 1uL of 10 fmol pDEST_R4_R2

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