**2.7 PCR amplification**

*Biotechnological Applications of Biomass*

**514**

**Primer name** Membrane Protein (*MEP*)

Elongation Factor 1 alpha (*EF1α*)

*β*-TubM

**Table 1.**

*Specific primers used in the current study.*

**Forward primer (5′-3′)**

TGTACTCGGCAATGCTCTTG

CGTTTCTGCCCTCTCCCA

TCTTCATGGTTGGCTTCGCT

**Reverse primer (5′-3′)**

TTTGATGCTCCAGGCTTACC

TGCTTGACACGTGACGATGA

CTTGGGTCGAACATCTGCT

**Product size (bp)**

203 102 118

62 0 C

62 0 C

**Ta** 64 0 C

Manoli et al. [17]

Nicolaisen et al. [5]

Mitema et al. [18]

**Reference**

Conventional polymerase chain reaction amplification was carried out in volumes of 25 μl and consisted of 0.5 μl of 10 μM dNTPs (Bioline), 10× reaction buffer with MgCl2, 1 μl of 10 μM forward and reverse primers, 0.2 μl Kapa Taq, 1 μl of 10 ng DNA template, and sterile H20. Protocol performed and followed for cycling conditions were: 1 cycle for 5 min at 94°C followed by 35× (for 30 s at 94°C, for 45 s at 60°C, for 90 s at 72°C). Elongation step was achieved at 72°C for 7 min and finally at 4°C for 1 min. The products of PCR were assessed on 2% agarose/EtBr gel in TAE1 X buffer (Tris–acetate 40 mM and EDTA 1.0 mM). Fermentas (100 bp DNA ladder) was used as a molecular size marker.
