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A: This answer depends on whether you are pursuing the $1 million direct investment or $500,000 regional investment center route.
Direct Investment: For those persons making the direct investment, USCIS effectively requires that the entire capital amount be invested and at risk in the commercial enterprise at the time the first stage of the case is filed. Often times, in order to reduce the risk of making such a large investment and not being granted a green card afterwards, persons place funds into escrow (third party trust) accounts with the transfer of funds being made contingent on the approval of the first stage of the green card case (I-526).
Regional Investment Center: For those persons who decide to make at least a $500,000 investment into their chosen investment center, most centers place all funds into an escrow (third party trust) account pending the approval of the first stage of the green card case (I-526), after which time the funds are transferred.