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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:
| DNA | DNA 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) | |
| l | x | y | f | 0.66*y*l*f/x | |||
| pENTR_LilrB2 | 4.4 | 200 | 100 | 5 | 7.26 | 92.74 | |
| pENTR_PirB | 5.1 | 200 | 100 | 5 | 8.415 | 91.585 | |
| pENTR_Hef1a | 3.9 | 200 | 100 | 5 | 6.435 | 93.565 | |
| pDEST | 7.9 | 200 | 100 | 10 | 26.07 | 73.93 |
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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