Table

Table 2. Primer combinations for PCR and sequencing

Rescued end PCR primers Annealing temperature Sequencing primersa
P-element Forward Reverse
P{Z} 5′ Plac4 Plac1 60ºC Splac2 Sp1
P{Z} 3′ Pry4 Pry1 55ºC Spep1 Sp3
P{lacW} 5′ Plac4 Plac1 60ºC Splac2 Sp1
P{lacW}b 3′ Pry4 Plw3-1 55ºC Spep1 Sp6(Sau3AI)
Sp5(HinP1I)
P{EP} 5′ Pwht1 Plac1 60ºC Sp1 --
P{EP} 3′ Pry4 Pry1 55ºC Spep1 --
aWhere two primers are listed, the inverse PCR product can be sequenced from both ends by setting up sequencing reactions with the two different primers.
bFor the rescue of sequences 3′ of P{lacW}, use the Sp6 primer on templates derived from Sau3AI digests, and Sp5 primer on templates derived from HinP1I digests. The Spep1 primer works on both templates.

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  1. doi:10.1101/pdb.tab295199 Cold Spring Harb Protoc 2009: pdb.tab295199-

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